Biodegradable lipids for delivery of nucleic acids

ABSTRACT

The present invention provides, in part, a biodegradable compound of formula I, and sub-formulas thereof: Formula (I) or a pharmaceutically acceptable salt thereof, where each X independently is O or S, each Y independently is O or S, and each R 1  independently is defined herein; and a liposome composition comprising the cationic lipid of formula I or a sub-formula thereof, and methods of delivering agents, such as nucleic acids including mRNA, in vivo, by administering to a subject the liposome comprising the cationic lipid of formula I or a sub-formula thereof, where the agent is encapsulated within the liposome.

BACKGROUND

The delivery of agents, such as nucleic acids, has been explored extensively as a potential therapeutic option for certain disease states. In particular, RNA interference (RNAi) has been the subject of significant research and clinical development. Lately, messenger RNA (mRNA) therapy has become an increasingly important option for treatment of various diseases, in particular, for those associated with deficiency of one or more proteins.

SUMMARY OF THE INVENTION

The present invention provides, among other things, a novel class of lipid compounds for improved in vivo delivery of therapeutic agents, such as nucleic acids. In particular, the compounds provided by the present invention are biodegradable in nature and are particularly useful for delivery of mRNA and other nucleic acids for therapeutic uses. It is contemplated that the compounds provided herein are capable of highly effective in vivo delivery while maintaining favorable toxicity profile due to the biodegradable nature.

In one aspect, the present invention provides a compound (i.e., cationic lipid) of formula I:

or a pharmaceutically acceptable salt thereof,

wherein:

each X independently is O or S;

each Y independently is O or S; and

each R¹ independently is defined herein.

In some embodiments, the compound of formula I is of formula II:

or a pharmaceutically acceptable salt thereof, wherein:

each X independently is O or S;

each Y independently is O or S;

each m independently is 0 to 20;

each n independently is 1 to 6;

each R_(A) is independently hydrogen, optionally substituted C1-50 alkyl, optionally substituted C2-50 alkenyl, optionally substituted C2-50 alkynyl, optionally substituted C3-10 carbocyclyl, optionally substituted 3-14 membered heterocyclyl, optionally substituted C6-14 aryl, optionally substituted 5-14 membered heteroaryl or halogen, and

each R_(B) is independently hydrogen, optionally substituted C1-50 alkyl, optionally substituted C2-50 alkenyl, optionally substituted C2-50 alkynyl, optionally substituted C3-10 carbocyclyl, optionally substituted 3-14 membered heterocyclyl, optionally substituted C6-14 aryl, optionally substituted 5-14 membered heteroaryl or halogen.

In some embodiments, the compound has a structure of formula III (i.e., Target 23 or T23):

or a pharmaceutically acceptable salt thereof.

In some embodiments, a compound of formula II is a compound of formula IV (i.e., Target 24 or T24):

or a pharmaceutically acceptable salt thereof.

In some embodiments, a compound of formula II is a compound of formula V:

or a pharmaceutically acceptable salt thereof.

In another aspect, the invention provides a composition, such as a lipid nanoparticle (e.g., liposome), comprising one or more of the compounds (i.e., cationic lipids) of formula I, formula II, formula III, formula IV, formula V or a sub-formula thereof.

In some embodiments, a suitable composition of the present invention is a liposome. In some embodiments, a suitable liposome comprises one or more cationic lipids of formula I, formula II, formula III, formula IV, formula V or a sub-formula thereof. In particular embodiments, a suitable liposome comprises a cationic lipid of formula III. In particular embodiments, a suitable liposome comprises a cationic lipid of formula IV. In particular embodiments, a suitable liposome comprises a cationic lipid of formula V.

In some embodiments, a suitable liposome further comprises one or more non-cationic lipids, one or more cholesterol-based lipids and/or one or more PEG-modified lipids. In some embodiments, the one or more non-cationic lipids are selected from distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoyl-phosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidyl-glycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidyl-choline (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE), dioleoylphosphatidyl-ethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, 1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), or a mixture thereof.

In some embodiments, a suitable liposome further comprises one or more cholesterol-based lipids. In some embodiments, the one or more cholesterol-based lipids are selected from cholesterol, PEGylated cholesterol and DC-Chol (N,N-dimethyl-N-ethylcarbox-amidocholesterol), 1,4-bis(3-N-oleylamino-propyl)piperazine.

In some embodiments, a suitable liposome further comprises one or more PEG-modified lipids. In some embodiments, the one or more PEG-modified lipids comprise a poly(ethylene) glycol chain of up to 5 kDa in length covalently attached to a lipid with alkyl chain(s) of C₆-C₂₀ length. In some embodiments, a PEG-modified lipid is a derivatized ceramide such as N-Octanoyl-Sphingosine-1-[Succinyl(Methoxy Polyethylene Glycol)-2000]. In some embodiments, a PEG-modified or PEGylated lipid is PEGylated cholesterol or Dimyristoylglycerol (DMG)-PEG-2K.

In some embodiments, a suitable liposome comprises the compound of Formula III, DOPE, cholesterol and DMG-PEG2K.

In some embodiments, a suitable liposome has a size of or less than about 500 nm, 450 nm, 400 nm, 350 nm, 300 nm, 250 nm, 200 nm, 150 nm, 125 nm, 110 nm, 100 nm, 95 nm, 90 nm, 85 nm, 80 nm, 75 nm, 70 nm, 65 nm, 60 nm, 55 nm, or 50 nm.

In some embodiments, a liposome according to the present invention comprises an mRNA encoding a protein encapsulated therein.

In yet another aspect, the present invention provides methods of delivering a therapeutic agent, such as a nucleic acid (e.g., DNA, siRNA, mRNA, microRNA) using a composition (e.g., liposome) described herein. In still another aspect, the present invention provides methods of treating a disease or disorder including administering to subject in need of treatment a composition (e.g., liposome) comprising a therapeutic agent, such as a nucleic acid (e.g., DNA, siRNA, mRNA, microRNA).

Other features, objects, and advantages of the present invention are apparent in the detailed description, drawings and claims that follow. It should be understood, however, that the detailed description, the drawings, and the claims, while indicating embodiments of the present invention, are given by way of illustration only, not limitation. Various changes and modifications within the scope of the invention will become apparent to those skilled in the art.

BRIEF DESCRIPTION OF THE DRAWING

Illustrated in FIG. 1 are human EPO levels in wild type mouse sera after treatment via hEPO mRNA loaded LNPs. Treatment after 6 hours is shown in the bars at right. Treatment after 24 hours is shown in the bars at left.

DEFINITIONS

In order for the present invention to be more readily understood, certain terms are first defined below. Additional definitions for the following terms and other terms are set forth throughout the specification. The publications and other reference materials referenced herein to describe the background of the invention and to provide additional detail regarding its practice are hereby incorporated by reference.

Chemical Definitions

Definitions of specific functional groups and chemical terms are described in more detail below. The chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed., inside cover, and specific functional groups are generally defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999; Smith and March March's Advanced Organic Chemistry, 5th Edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; and Carruthers, Some Modern Methods of Organic Synthesis, 3rd Edition, Cambridge University Press, Cambridge, 1987.

Compounds described herein can comprise one or more asymmetric centers, and thus can exist in various isomeric forms, e.g., enantiomers and/or diastereomers. For example, the compounds described herein can be in the form of an individual enantiomer, diastereomer or geometric isomer, or can be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomer. Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric syntheses. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, E. L. Stereochemistry of Carbon Compounds (McGraw-Hill, N Y, 1962); and Wilen, S. H. Tables of Resolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind. 1972). The invention additionally contemplates compounds as individual isomers substantially free of other isomers, and alternatively, as mixtures of various isomers.

When a range of values is listed, it is intended to encompass each value and sub-range within the range. For example “C1-6 alkyl” is intended to encompass, C1, C2, C3, C4, C5, C6, C1-6, C1-5, C1-4, C1-3, C1-2, C2-6, C2-5, C2-4, C2-3, C3-6, C3-5, C3-4, C4-6, C4-5, and C5-6 alkyl.

As used herein, “alkyl” refers to a radical of a straight-chain or branched saturated hydrocarbon group having from 1 to 50 carbon atoms (“C1-50 alkyl”). In some embodiments, an alkyl group has 1 to 40 carbon atoms (“C1-40 alkyl”). In some embodiments, an alkyl group has 1 to 30 carbon atoms (“C1-30 alkyl”). In some embodiments, an alkyl group has 1 to 20 carbon atoms (“C1-20 alkyl”). In some embodiments, an alkyl group has 1 to 10 carbon atoms (“C1-10 alkyl”). In some embodiments, an alkyl group has 1 to 9 carbon atoms (“C1-9 alkyl”). In some embodiments, an alkyl group has 1 to 8 carbon atoms (“C1-8 alkyl”). In some embodiments, an alkyl group has 1 to 7 carbon atoms (“C1-7 alkyl”). In some embodiments, an alkyl group has 1 to 6 carbon atoms (“C1-6 alkyl”). In some embodiments, an alkyl group has 1 to 5 carbon atoms (“C1-5 alkyl”). In some embodiments, an alkyl group has 1 to 4 carbon atoms (“C1-4 alkyl”). In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C1-3 alkyl”). In some embodiments, an alkyl group has 1 to 2 carbon atoms (“C1-2 alkyl”). In some embodiments, an alkyl group has 1 carbon atom (“C1 alkyl”). In some embodiments, an alkyl group has 2 to 6 carbon atoms (“C2-6 alkyl”). Examples of C1-6 alkyl groups include, without limitation, methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), iso-butyl (C4), n-pentyl (C5), 3-pentanyl (C5), amyl (C5), neopentyl (C5), 3-methyl-2-butanyl (C5), tertiary amyl (C5), and n-hexyl (C6). Additional examples of alkyl groups include n-heptyl (C7), n-octyl (C8) and the like. Unless otherwise specified, each instance of an alkyl group is independently unsubstituted (an “unsubstituted alkyl”) or substituted (a “substituted alkyl”) with one or more substituents. In certain embodiments, the alkyl group is an unsubstituted C1-50 alkyl. In certain embodiments, the alkyl group is a substituted C1-50 alkyl.

As used herein, “heteroalkyl” refers to an alkyl group as defined herein which further includes at least one heteroatom (e.g., 1 to 25, e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus within (i.e., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain. In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 50 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-50 alkyl”). In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 40 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-40 alkyl”). In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 30 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-30 alkyl”). In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 20 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-20 alkyl”). In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 10 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-10 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 9 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-9 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 8 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-8 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 7 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-7 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 6 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-6 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 5 carbon atoms and 1 or 2 heteroatoms within the parent chain (“heteroC1-5 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 4 carbon atoms and 1 or 2 heteroatoms within the parent chain (“heteroC1-4 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 3 carbon atoms and 1 heteroatom within the parent chain (“heteroC1-3 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 2 carbon atoms and 1 heteroatom within the parent chain (“heteroC1-2 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 carbon atom and 1 heteroatom (“heteroC1 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 2 to 6 carbon atoms and 1 or 2 heteroatoms within the parent chain (“heteroC2-6 alkyl”). Unless otherwise specified, each instance of a heteroalkyl group is independently unsubstituted (an “unsubstituted heteroalkyl”) or substituted (a “substituted heteroalkyl”) with one or more substituents. In certain embodiments, the heteroalkyl group is an unsubstituted heteroC1-50 alkyl. In certain embodiments, the heteroalkyl group is a substituted heteroC1-50 alkyl.

As used herein, “alkenyl” refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 50 carbon atoms and one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 double bonds) (“C2-50 alkenyl”). In some embodiments, an alkenyl group has 2 to 40 carbon atoms (“C2-40 alkenyl”). In some embodiments, an alkenyl group has 2 to 30 carbon atoms (“C2-30 alkenyl”). In some embodiments, an alkenyl group has 2 to 20 carbon atoms (“C2-20 alkenyl”). In some embodiments, an alkenyl group has 2 to 10 carbon atoms (“C2-10 alkenyl”). In some embodiments, an alkenyl group has 2 to 9 carbon atoms (“C2-9 alkenyl”). In some embodiments, an alkenyl group has 2 to 8 carbon atoms (“C2-8 alkenyl”). In some embodiments, an alkenyl group has 2 to 7 carbon atoms (“C2-7 alkenyl”). In some embodiments, an alkenyl group has 2 to 6 carbon atoms (“C2-6 alkenyl”). In some embodiments, an alkenyl group has 2 to 5 carbon atoms (“C2-5 alkenyl”). In some embodiments, an alkenyl group has 2 to 4 carbon atoms (“C2-4 alkenyl”). In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C2-3 alkenyl”). In some embodiments, an alkenyl group has 2 carbon atoms (“C2 alkenyl”). The one or more carbon-carbon double bonds can be internal (such as in 2-butenyl) or terminal (such as in 1-butenyl). Examples of C2-4 alkenyl groups include, without limitation, ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), and the like. Examples of C2-6 alkenyl groups include the aforementioned C2-4 alkenyl groups as well as pentenyl (C5), pentadienyl (C5), hexenyl (C6), and the like. Additional examples of alkenyl include heptenyl (C7), octenyl (C8), octatrienyl (C8), and the like. Unless otherwise specified, each instance of an alkenyl group is independently unsubstituted (an “unsubstituted alkenyl”) or substituted (a “substituted alkenyl”) with one or more substituents. In certain embodiments, the alkenyl group is an unsubstituted C2-50 alkenyl. In certain embodiments, the alkenyl group is a substituted C2-50 alkenyl.

As used herein, “heteroalkenyl” refers to an alkenyl group as defined herein which further includes at least one heteroatom (e.g., 1 to 25, e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus within (i.e., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain. In certain embodiments, a heteroalkenyl group refers to a group having from 2 to 50 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-50 alkenyl”). In certain embodiments, a heteroalkenyl group refers to a group having from 2 to 40 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-40 alkenyl”). In certain embodiments, a heteroalkenyl group refers to a group having from 2 to 30 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-30 alkenyl”). In certain embodiments, a heteroalkenyl group refers to a group having from 2 to 20 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-20 alkenyl”). In certain embodiments, a heteroalkenyl group refers to a group having from 2 to 10 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-10 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 9 carbon atoms at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-9 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 8 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-8 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 7 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-7 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 6 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-6 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 5 carbon atoms, at least one double bond, and 1 or 2 heteroatoms within the parent chain (“heteroC2-5 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 4 carbon atoms. at least one double bond, and for 2 heteroatoms within the parent chain (“heteroC2-4 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 3 carbon atoms, at least one double bond, and 1 heteroatom within the parent chain (“heteroC2-3 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 6 carbon atoms, at least one double, bond, and 1 or 2 heteroatoms within the parent chain (“heteroC2-6 alkenyl”). Unless otherwise specified, each instance of a heteroalkenyl group is independently unsubstituted (an “unsubstituted heteroalkenyl”) or substituted (a “substituted heteroalkenyl”) with one or more substituents. In certain embodiments, the heteroalkenyl group is an unsubstituted heteroC2-50 alkenyl. In certain embodiments, the heteroalkenyl group is a substituted heteroC2-50 alkenyl.

As used herein, “alkynyl” refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 50 carbon atoms and one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 triple bonds) and optionally one or more double bonds (e.g., 1, 2, 3, or 4 double bonds) (“C2-50 alkynyl”). An alkynyl group that has one or more triple bonds and one or more double bonds is also referred to as an “ene-yne”. In some embodiments, an alkynyl group has 2 to 40 carbon atoms (“C2-40 alkynyl”). In some embodiments, an alkynyl group has 2 to 30 carbon atoms (“C2-30 alkynyl”). In some embodiments, an alkynyl group has 2 to 20 carbon atoms (“C2-20 alkynyl”). In some embodiments, an alkynyl group has 2 to 10 carbon atoms (“C2-10 alkynyl”). In some embodiments, an alkynyl group has 2 to 9 carbon atoms (“C2-9 alkynyl”). In some embodiments, an alkynyl group has 2 to 8 carbon atoms (“C2-8 alkynyl”). In some embodiments, an alkynyl group has 2 to 7 carbon atoms (“C2-7 alkynyl”). In some embodiments, an alkynyl group has 2 to 6 carbon atoms (“C2-6 alkynyl”). In some embodiments, an alkynyl group has 2 to 5 carbon atoms (“C2-5 alkynyl”). In some embodiments, an alkynyl group has 2 to 4 carbon atoms (“C2-4 alkynyl”). In some embodiments, an alkynyl group has 2 to 3 carbon atoms (“C2-3 alkynyl”). In some embodiments, an alkynyl group has 2 carbon atoms (“C2 alkynyl”). The one or more carbon-carbon triple bonds can be internal (such as in 2-butynyl) or terminal (such as in 1-butynyl). Examples of C2-4 alkynyl groups include, without limitation, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), and the like. Examples of C2-6 alkenyl groups include the aforementioned C2-4 alkynyl groups as well as pentynyl (C5), hexynyl (C6), and the like. Additional examples of alkynyl include heptynyl (C7), octynyl (C8), and the like. Unless otherwise specified, each instance of an alkynyl group is independently unsubstituted (an “unsubstituted alkynyl”) or substituted (a “substituted alkynyl”) with one or more substituents. In certain embodiments, the alkynyl group is an unsubstituted C2-50 alkynyl. In certain embodiments, the alkynyl group is a substituted C2-50 alkynyl.

As used herein, “heteroalkynyl” refers to an alkynyl group as defined herein which further includes at least one heteroatom (e.g., 1 to 25, e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus within (i.e., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain. In certain embodiments, a heteroalkynyl group refers to a group having from 2 to 50 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-50 alkynyl”). In certain embodiments, a heteroalkynyl group refers to a group having from 2 to 40 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-40 alkynyl”). In certain embodiments, a heteroalkynyl group refers to a group having from 2 to 30 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-30 alkynyl”). In certain embodiments, a heteroalkynyl group refers to a group having from 2 to 20 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-20 alkynyl”). In certain embodiments, a heteroalkynyl group refers to a group having from 2 to 10 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-10 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 9 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-9 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 8 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-8 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 7 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-7 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 6 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-6 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 5 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms within the parent chain (“heteroC2-5 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 4 carbon atoms, at least one triple bond, and for 2 heteroatoms within the parent chain (“heteroC2-4 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 3 carbon atoms, at least one triple bond, and 1 heteroatom within the parent chain (“heteroC2-3 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 6 carbon atoms, at least one triple bond. and 1 or 2 heteroatoms within the parent chain (“heteroC2-6 alkynyl”). Unless otherwise specified, each instance of a heteroalkynyl group is independently unsubstituted (an “unsubstituted heteroalkynyl”) or substituted (a “substituted heteroalkynyl”) with one or more substituents. In certain embodiments, the heteroalkynyl group is an unsubstituted heteroC2-50 alkynyl. In certain embodiments, the heteroalkynyl group is a substituted heteroC2-50 alkynyl.

As used herein, “carbocyclyl” or “carbocyclic” refers to a radical of a non-aromatic cyclic hydrocarbon group having from 3 to 10 ring carbon atoms (“C3-10 carbocyclyl”) and zero heteroatoms in the non-aromatic ring system. In some embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms (“C3-8 carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to 7 ring carbon atoms (“C3-7 carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms (“C3-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 4 to 6 ring carbon atoms (“C4-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to 6 ring carbon atoms (“C5-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms (“C5-10 carbocyclyl”). Exemplary C3-6 carbocyclyl groups include, without limitation, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), and the like. Exemplary C3-8 carbocyclyl groups include, without limitation, the aforementioned C3-6 carbocyclyl groups as well as cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo[2.2.1]heptanyl (C7), bicyclo[2.2.2]octanyl (C8), and the like. Exemplary C3-10 carbocyclyl groups include, without limitation, the aforementioned C3-8 carbocyclyl groups as well as cyclononyl (C9), cyclononenyl (C9), cyclodecyl (C10), cyclodecenyl (C10), octahydro-1H-indenyl (C9), decahydronaphthalenyl (C10), spiro[4.5]decanyl (C10), and the like. As the foregoing examples illustrate, in certain embodiments, the carbocyclyl group is either monocyclic (“monocyclic carbocyclyl”) or polycyclic (e.g., containing a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic carbocyclyl”) or tricyclic system (“tricyclic carbocyclyl”)) and can be saturated or can contain one or more carbon-carbon double or triple bonds. “Carbocyclyl” also includes ring systems wherein the carbocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups wherein the point of attachment is on the carbocyclyl ring, and in such instances, the number of carbons continue to designate the number of carbons in the carbocyclic ring system. Unless otherwise specified, each instance of a carbocyclyl group is independently unsubstituted (an “unsubstituted carbocyclyl”) or substituted (a “substituted carbocyclyl”) with one or more substituents. In certain embodiments, the carbocyclyl group is an unsubstituted C3-10 carbocyclyl. In certain embodiments, the carbocyclyl group is a substituted C3-10 carbocyclyl.

In some embodiments, “carbocyclyl” or “carbocyclic” is referred to as a “cycloalkyl”, i.e., a monocyclic, saturated carbocyclyl group having from 3 to 10 ring carbon atoms (“C3-10 cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms (“C3-8 cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms (“C3-6, cycloalkyl”). In some embodiments, a cycloalkyl group has 4 to 6 ring carbon atoms (“C4-6 cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C5-6 cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms (“C5-10 cycloalkyl”). Examples of C5-6 cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (C5). Examples of C3-6 cycloalkyl groups include the aforementioned C5-6 cycloalkyl groups as well as cyclopropyl (C3) and cyclobutyl (C4). Examples of C3-8 cycloalkyl groups include the aforementioned C3-6 cycloalkyl groups as well as cycloheptyl (C7) and cyclooctyl (C8). Unless otherwise specified, each instance of a cycloalkyl group is independently unsubstituted (an “unsubstituted cycloalkyl”) or substituted (a “substituted cycloalkyl”) with one or more substituents. In certain embodiments, the cycloalkyl group is an unsubstituted C3-10 cycloalkyl. In certain embodiments, the cycloalkyl group is a substituted C3-10 cycloalkyl.

As used herein, “heterocyclyl” or “heterocyclic” refers to a radical of a 3- to 14-membered non-aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“3-14 membered heterocyclyl”). In heterocyclyl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. A heterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”) or polycyclic (e.g., a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic heterocyclyl”) or tricyclic system (“tricyclic heterocyclyl”)). and can be saturated or can contain one or more carbon-carbon double or triple bonds. Heterocyclyl polycyclic ring systems can include one or more heteroatoms in one or both rings. “Heterocyclyl” also includes ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more carbocyclyl groups wherein the point of attachment is either on the carbocyclyl or heterocyclyl ring, or ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heterocyclyl ring system. Unless otherwise specified, each instance of heterocyclyl is independently unsubstituted (an “unsubstituted heterocyclyl”) or substituted (a “substituted heterocyclyl”) with one or more substituents. In certain embodiments, the heterocyclyl group is an unsubstituted 3-14 membered heterocyclyl. In certain embodiments, the heterocyclyl group is a substituted 3-14 membered heterocyclyl.

In some embodiments, a heterocyclyl group is a 5-10 membered non-aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-10 membered heterocyclyl”). In some embodiments, a heterocyclyl group is a 5-8 membered non-aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-8 membered heterocyclyl”). In some embodiments, a heterocyclyl group is a 5-6 membered non-aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-6 membered heterocyclyl”). In some embodiments, the 5-6 membered heterocyclyl has 1 or more (e.g., 1, 2, or 3) ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heterocyclyl has 1 or 2 ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heterocyclyl has 1 ring heteroatom selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus.

Exemplary 3-membered heterocyclyl groups containing 1 heteroatom include, without limitation, azirdinyl, oxiranyl, thiorenyl. Exemplary 4-membered heterocyclyl groups containing 1 heteroatom include, without limitation, azetidinyl, oxetanyl and thietanyl. Exemplary 5-membered heterocyclyl groups containing 1 heteroatom include, without limitation. tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclyl groups containing 2 heteroatoms include, without limitation, dioxolanyl, oxathiolanyl and dithiolanyl. Exemplary 5-membered heterocyclyl groups containing 3 heteroatoms include, without limitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclyl groups containing 1 heteroatom include, without limitation, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclyl groups containing 2 heteroatoms include, without limitation, piperazinyl, morpholinyl, dithianyl, dioxanyl. Exemplary 6-membered heterocyclyl groups containing 2 heteroatoms include, without limitation, triazinanyl. Exemplary 7-membered heterocyclyl groups containing 1 heteroatom include, without limitation, azepanyl, oxepanyl and thiepanyl. Exemplary 8-membered heterocyclyl groups containing 1 heteroatom include, without limitation, azocanyl, oxecanyl and thiocanyl. Exemplary bicyclic heterocyclyl groups include, without limitation, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, tetrahydrobenzothienyl, tetrahydrobenzofuranyl, tetrahydroindolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, decahydroisoquinolinyl, octahydrochromenyl, octahydroisochromenyl, decahydronaphthyridinyl, decahydro-1,8-naphthyridinyl, octahydropyrrolo[3,2-b]pyrrole, indolinyl, phthalimidyl, naphthalimidyl, chromanyl, chromenyl, 1H-benzo[e][1,4]diazepinyl, 1,4,5,7-tetrahydropyrano[3,4-b] pyrrolyl, 5,6-dihydro-4H-furo[3,2-b]pyrrolyl, 6,7-dihydro-5H-furo[3,2-b]pyranyl, 5,7-dihydro-4H-thieno[2,3-c]pyranyl, 2,3-dihydro-1H-pyrrolo[2,3-b]pyridinyl, 2,3-dihydrofuro[2,3-b]pyridinyl, 4,5,6,7-tetrahydro-1H-pyrrolo-[2,3-b]pyridinyl, 4,5,6,7-tetrahydrofuro[3,2-c]pyridinyl, 4,5,6,7-tetrahydrothieno [3,2-b]pyridinyl, 1,2,3,4-tetrahydro-1,6-naphthyridinyl, and the like.

As used herein, “aryl” refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 nt electrons shared in a cyclic array) having 6-14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system (“C6-14 aryl”). In some embodiments, an aryl group has 6 ring carbon atoms (“C6 aryl”; e.g., phenyl). In some embodiments, an aryl group has 10 ring carbon atoms (“C10 aryl”; e.g., naphthyl such as 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has 14 ring carbon atoms (“C14 aryl”; e.g., anthracyl). “Aryl” also includes ring systems wherein the aryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the radical or point of attachment is on the aryl ring, and in such instances, the number of carbon atoms continue to designate the number of carbon atoms in the aryl ring system. Unless otherwise specified, each instance of an aryl group is independently unsubstituted (an “unsubstituted aryl”) or substituted (a “substituted aryl”) with one or more substituents. In certain embodiments, the aryl group is an unsubstituted C6-14 aryl. In certain embodiments, the aryl group is a substituted C6-14 aryl.

As used herein, “heteroaryl” refers to a radical of a 5-14 membered monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 it electrons shared in a cyclic array) having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4 ring heteroatoms) ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-14 membered heteroaryl”). In heteroaryl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. Heteroaryl polycyclic ring systems can include one or more heteroatoms in one or both rings. “Heteroaryl” includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the point of attachment is on the heteroaryl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heteroaryl ring system. “Heteroaryl” also includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more aryl groups wherein the point of attachment is either on the aryl or heteroaryl ring, and in such instances, the number of ring members designates the number of ring members in the fused polycyclic (aryl/heteroaryl) ring system. Polycyclic heteroaryl groups wherein one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and the like) the point of attachment can be on either ring, i.e., either the ring bearing a heteroatom (e.g., 2-indolyl) or the ring that does not contain a heteroatom (e.g., 5-indolyl).

In some embodiments, a heteroaryl group is a 5-10 membered aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-10 membered heteroaryl”). In some embodiments, a heteroaryl group is a 5-8 membered aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-8 membered heteroaryl”). In some embodiments, a heteroaryl group is a 5-6 membered aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-6 membered heteroaryl”). In some embodiments, the 5-6 membered heteroaryl has 1 or more (e.g., 1, 2, or 3) ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heteroaryl has 1 or 2 ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. Unless otherwise specified, each instance of a heteroaryl group is independently unsubstituted (an “unsubstituted heteroaryl”) or substituted (a “substituted heteroaryl”) with one or more substituents. In certain embodiments, the heteroaryl group is an unsubstituted 5-14 membered heteroaryl. In certain embodiments, the heteroaryl group is a substituted 5-14 membered heteroaryl.

Exemplary 5-membered heteroaryl groups containing 1 heteroatom include, without limitation, pyrrolyl, furanyl and thiophenyl. Exemplary 5-membered heteroaryl groups containing 2 heteroatoms include, without limitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing 3 heteroatoms include, without limitation, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing 4 heteroatoms include, without limitation, tetrazolyl. Exemplary 6-membered heteroaryl groups containing 1 heteroatom include, without limitation. pyridinyl. Exemplary 6-membered heteroaryl groups containing 2 heteroatoms include, without limitation, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing 3 or 4 heteroatoms include, without limitation, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing 1 heteroatom include, without limitation, azepinyl, oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groups include, without limitation, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, without limitation, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. Exemplary tricyclic heteroaryl groups include, without limitation, phenanthridinyl, dibenzofuranyl, carbazolyl, acridinyl, phenothiazinyl, phenoxazinyl and phenazinyl.

As used herein, the term “partially unsaturated” refers to a ring moiety that includes at least one double or triple bond. The term “partially unsaturated” is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aromatic groups (e.g., aryl or heteroaryl moieties) as herein defined.

As used herein, the term “saturated” refers to a ring moiety that does not contain a double or triple bond, i.e., the ring contains all single bonds.

Affixing the suffix “-ene” to a group indicates the group is a divalent moiety, e.g., alkylene is the divalent moiety of alkyl, alkenylene is the divalent moiety of alkenyl,

alkynylene is the divalent moiety of alkynyl, heteroalkylene is the divalent moiety of heteroalkyl, heteroalkenylene is the divalent moiety of heteroalkenyl, heteroalkynylene is the divalent moiety of heteroalkynyl, carbocyclylene is the divalent moiety of carbocyclyl, heterocyclylene is the divalent moiety of heterocyclyl, arylene is the divalent moiety of aryl, and heteroarylene is the divalent moiety of heteroaryl.

As understood from the above, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl groups, as defined herein, are, in certain embodiments, optionally substituted. Optionally substituted refers to a group which may be substituted or unsubstituted (e.g., “substituted” or “unsubstituted” alkyl, “substituted” or “unsubstituted” alkenyl, “substituted” or “unsubstituted” alkynyl, “substituted” or “unsubstituted” heteroalkyl, “substituted” or “unsubstituted” heteroalkenyl, “substituted” or “unsubstituted” heteroalkynyl. “substituted” or “unsubstituted” carbocyclyl. “substituted” or “unsubstituted” heterocyclyl, “substituted” or “unsubstituted” aryl or “substituted” or “unsubstituted” heteroaryl group). In general, the term “substituted” means that at least one hydrogen present on a group is replaced with a permissible substituent, e.g., a substituent which upon substitution results in a stable compound, e.g., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, or other reaction. Unless otherwise indicated, a “substituted” group has a substituent at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituent is either the same or different at each position. The term “substituted” is contemplated to include substitution with all permissible substituents of organic compounds, any of the substituents described herein that results in the formation of a stable compound. The present invention contemplates any and all such combinations in order to arrive at a stable compound. For purposes of this invention, heteroatoms such as nitrogen may have hydrogen substituents and/or any suitable substituent as described herein which satisfy the valencies of the heteroatoms and results in the formation of a stable moiety.

Exemplary carbon atom substituents include, but are not limited to, halogen, —CN, —NO2, —N3, —SO2H, —SO3H, —OH, —ORaa, —ON(Rbb)2, —N(Rbb)2, —N(Rbb)3+X—, —N(ORcc)Rbb, —SeH, —SeRaa, —SH, —SRaa, —SSRcc, —C(═O)Raa, —CO2H, —CHO, —C(ORcc)2, —CO2Raa, —OC(═O)Raa, —OCO2Raa, —C(═O)N(Rbb)2, —OC(═O)N(Rbb)2, —NRbbC(═O)Raa, —NRbbCO2Raa, —NRbbC(═O)N(Rbb)2, —C(═NRbb)Raa, —C(═NRbb)ORaa, —OC(═NRbb)Raa, —OC(═NRbb)ORaa, —C(═NRbb)N(Rbb)2, —OC(═NRbb)N(Rbb)2, —NRbbC(═NRbb)N(Rbb)2, —C(═O)NRbbSO2Raa, —NRbbSO2Raa, —SO2N(Rbb)2, —SO2Raa, —SO2ORaa, —OSO2Raa, —S(═O)Raa, —OS(═O)Raa, —Si(Raa)3-OSi(Raa)3-C(═S)N(Rbb)2, —C(═O)SRaa, —C(═S)SRaa, —SC(═S)SRaa, —SC(═O)SRaa, —OC(═O)SRaa, —SC(═O)ORaa, —SC(═O)Raa, —P(═O)2Raa, —OP(═O)2Raa, —P(═O)(Raa)2, —OP(═O)(Raa)2, —OP(═O)(ORcc)2, —P(═O)2N(Rbb)2, —OP(═O)2N(Rbb)2, —P(═O)(NRbb)2, —OP(═O)(NRbb)2, —NRbbP(═O)(ORcc)2, —NRbbP(═O)(NRbb)2, —P(Rcc)2, —P(Rcc)3, —OP(Rcc)2, —OP(Rcc)3, —B(Raa)2, —B(ORcc)2, —BRaa(ORcc), C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-14 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups;

or two geminal hydrogens on a carbon atom are replaced with the group ═O, ═S, ═NN(Rbb)2, =NNRbbC(═O)Raa, =NNRbbC(═O)ORaa, =NNRbbS(═O)2Raa, =NRbb, or =NORcc;

each instance of Raa is, independently, selected from C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Raa groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups;

each instance of Rbb is, independently, selected from hydrogen, —OH, —ORaa, —N(Rcc)2, —CN, —C(═O)Raa, —C(═O)N(Rcc)2, —CO2Raa, —SO2Raa, —C(═NRcc)ORaa, —C(═NRcc)N(Rcc)2, —SO2N(Rcc)2, —SO2Rcc, —SO20Rcc, —SORaa, —C(═S)N(Rcc)2, —C(═O)SRcc, —C(═S)SRcc, —P(═O)2Raa, —P(═O)(Raa)2, —P(═O)2N(Rcc)2, —P(═O)(NRcc)2, C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Rbb groups, together with the heteroatom to which they are attached, form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups;

each instance of Rcc is, independently, selected from hydrogen, C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Rcc groups, together with the heteroatom to which they are attached, form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups;

each instance of Rdd is, independently, selected from halogen, —CN, —N02, —N3, —SO2H, —SO3H, —OH, —ORee, —ON(Rff)2, —N(Rff)2, —N(Rff)3+X—, —N(ORee)Rff, —SH, —SRee, —SSRee, —C(═O)Ree, —CO2H, —CO2Ree, —OC(═O)Ree, —OCO2Ree, —C(═O)N(Rff)2, —OC(═O)N(Rff)2, —NRffC(═O)Ree, —NRffCO2Ree, —NRffC(═O)N(Rff)2, —C(═NRff)ORee, —OC(═NRff)Ree, —OC(═NRff)ORee, —C(═NRff)N(Rff)2, —OC(═NRff)N(Rff)2, —NRffC(═NRff)N(Rff)2, —NRffSO2Ree, —SO2N(Rff)2, —SO2Ree, —SO2ORee, —OSO2Ree, —S(═O)Ree, —Si(Ree)3, —OSi(Ree)3, —C(═S)N(Rff)2, —C(═O)SRee, —C(═S)SRee, —SC(═S)SRee, —P(═O)2Ree, —P(═O)(Ree)2, —OP(═O)(Ree)2, —OP(═O)(ORee)2, C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, 3-10 membered heterocyclyl, C6-10 aryl, 5-10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgg groups, or two geminal Rdd substituents can be joined to form ═O or ═S;

each instance of Ree is, independently, selected from C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, C6-10 aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgg groups;

each instance of Rff is, independently, selected from hydrogen, C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, 3-10 membered heterocyclyl, C6-10 aryl and 5-10 membered heteroaryl, or two Rff groups, together with the heteroatom to which they are attached, form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgg groups; and

each instance of Rgg is, independently, halogen, —CN, —NO2, —N3, —SO2H, —SO3H, —OH, —OC1-50 alkyl, —ON(C1-50 alkyl)2, —N(C1-50 alkyl)2, —N(C1-50 alkyl)3+X—, —NH(C1-50 alkyl)2+X—, —NH2(C1-50 alkyl)+X—, —NH3+X—, —N(OC1-50 alkyl)(C1-50 alkyl), —N(OH)(C1-50 alkyl), —NH(OH), —SH, —SC1-50 alkyl, —SS(C1-50 alkyl), —C(═O)(C1-50 alkyl), —CO2H, —CO2(C1-50 alkyl), —OC(═O)(C1-50 alkyl), —OCO2(C1-50 alkyl), —C(═O)NH2, —C(═O)N(C1-50 alkyl)2, —OC(═O)NH(C1-50 alkyl), —NHC(═O)(C1-50 alkyl), —N(C1-50 alkyl)C(═O)(C1-50 alkyl), —NHCO2(C1-50 alkyl), —NHC(═O)N(C1-50 alkyl)2, —NHC(═O)NH(C1-50 alkyl), —NHC(═O)NH2, —C(═NH)O(C1-50 alkyl), —OC(═NH)(C1-50 alkyl), —OC(═NH)OC1-50 alkyl, —C(═NH)N(C1-50 alkyl)2, —C(═NH)NH(C1-50 alkyl), —C(═NH)NH2, —OC(═NH)N(C1-50alkyl)2, —OC(NH)NH(C1-50 alkyl), —OC(NH)NH2, —NHC(NH)N(C1-50 alkyl)2, —NHC(═NH)NH2, —NHSO2 (C1-50 alkyl), —SO2N(C1-50 alkyl)2, —SO2NH(C1-50 alkyl), —SO2NH2, —SO2C1-50 alkyl, —SO2OC1-50 alkyl, —OSO2C1-6 alkyl, —SOC1-6 alkyl, —Si(C1-50 alkyl)3, —OSi(C1-6 alkyl)3-C(═S)N(C1-50 alkyl)2, C(═S)NH(C1-50 alkyl), C(═S)NH2, —C(═O)S(C1-6 alkyl), —C(═S)SC1-6 alkyl, —SC(═S)SC1-6 alkyl, —P(═O)2(C1-50 alkyl), —P(═O)(C1-50 alkyl)2, —OP(═O)(C1-50 alkyl)2, —OP(═O)(OC1-50 alkyl)2, C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, C6-10 aryl, 3-10 membered heterocyclyl, 5-10 membered heteroaryl; or two geminal Rgg substituents can be joined to form ═O or ═S; wherein X— is a counterion.

As used herein, the term “halo” or “halogen” refers to fluorine (fluoro, —F), chlorine (chloro, —Cl), bromine (bromo, —Br), or iodine (iodo, —I).

As used herein, a “counterion” is a negatively charged group associated with a positively charged quarternary amine in order to maintain electronic neutrality. Exemplary counterions include halide ions (e.g., F—, Cl—, Br—, I—), NO3-, C1O4-, OH—, H2PO4-, HSO4-, sulfonate ions (e.g., methansulfonate, trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate, 10-camphor sulfonate, naphthalene-2-sulfonate, naphthalene-1-sulfonic acid-5-sulfonate, ethan-1-sulfonic acid-2-sulfonate, and the like), and carboxylate ions (e.g., acetate, ethanoate, propanoate, benzoate, glycerate, lactate, tartrate, glycolate, and the like).

Nitrogen atoms can be substituted or unsubstituted as valency permits, and include primary, secondary, tertiary, and quarternary nitrogen atoms. Exemplary nitrogen atom substitutents include, but are not limited to, hydrogen, —OH, —ORaa, —N(Rcc)2, —CN, —C(═O)Raa, —C(═O)N(Rcc)2, —CO2Raa, —SO2Raa, —C(═NRbb)Raa, —C(═NRcc)ORaa, —C(═NRcc)N(Rcc)2, —SO2N(Rcc)2, —SO2Rcc, —SO2ORcc, —SORaa, —C(═S)N(Rcc)2, —C(═O)SRcc, —C(═S)SRcc, —P(═O)2Raa, —P(═O)(Raa)2, —P(═O)2N(Rcc)2, —P(═O)(NRcc)2, C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Rcc groups, together with the N atom to which they are attached, form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups, and wherein Raa, Rbb, Rcc and Rdd are as defined above.

Nitrogen atoms can be substituted or unsubstituted as valency permits, and include primary, secondary, tertiary, and quarternary nitrogen atoms. Exemplary nitrogen atom substitutents include, but are not limited to, hydrogen, —OH, —ORaa, —N(Rcc)2, —CN, —C(═O)Raa, —C(═O)N(Rcc)2, —CO2Raa, —SO2Raa, —C(═NRbb)Raa, —C(═NRcc)ORaa, —C(═NRcc)N(Rcc)2, —SO2N(Rcc)2, —SO2Rcc, —SO2ORcc, —SORaa, —C(═S)N(Rcc)2, —C(═O)SRcc, —C(═S)SRcc, —P(═O)2Raa, —P(═O)(Raa)2, —P(═O)2N(Rcc)2, —P(═O)(NRcc)2, C1-10 alkyl, C1-10 perhaloalkyl, C2-10 alkenyl, C2-10 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Rcc groups, together with the nitrogen atom to which they are attached, form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups, and wherein Raa, Rbb, Rcc and Rdd are as defined above.

In certain embodiments, the substituent present on a nitrogen atom is a nitrogen protecting group (also referred to as an amino protecting group). Nitrogen protecting groups include, but are not limited to, —OH, —ORaa, —N(Rcc)2, —C(═O)Raa, —C(═O)N(Rcc)2, —CO2Raa, —SO2Raa, —C(═NRcc)Raa, —C(═NRcc)ORaa, —C(═NRcc)N(Rcc)2, —SO2N(Rcc)2, —SO2Rcc, —SO2ORcc, —SORaa, —C(═S)N(Rcc)2, —C(═O)SRcc, —C(═S)SRcc, C1-10 alkyl (e.g., aralkyl, heteroaralkyl), C2-10 alkenyl, C2-10 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl groups, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aralkyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 Rdd groups, and wherein Raa, Rbb, Rcc and Rdd are as defined herein. Nitrogen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999, incorporated herein by reference.

For example, nitrogen protecting groups such as amide groups (e.g., —C(═O)Raa) include, but are not limited to, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivative, benzamide, p-phenylbenzamide, o-nitophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide, (N′-dithiobenzyloxyacylamino)acetamide, 3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine derivative, o-nitrobenzamide and o-(benzoyloxymethyl)benzamide.

Nitrogen protecting groups such as carbamate groups (e.g., —C(═O)ORaa) include, but are not limited to, methyl carbamate, ethyl carbamante, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc), 2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate (Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethyl carbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC), 1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc), 1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2′- and 4′-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithio carbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p-nitobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate, 2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methyl carbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc), 2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate (Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate, 2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methyl carbamate, t-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate, 2,2-dimethoxyacylvinyl carbamate, o-(N,N-dimethylcarboxamido)benzyl carbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate, 1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate, 2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl carbamate, isobutyl carbamate, isonicotinyl carbamate, p-(p′-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate, 1-methyl-1(3,5-dimethoxyphenyl)ethyl carbamate, 1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl carbamate, and 2,4,6-trimethylbenzyl carbamate.

Nitrogen protecting groups such as sulfonamide groups (e.g., —S(═O)2Raa) include, but are not limited to, p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms), f3-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide, 4-(4′,8′-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

Other nitrogen protecting groups include, but are not limited to, phenothiazinyl-(10)-acyl derivative, N′-p-toluenesulfonylaminoacyl derivative, N′-phenylaminothioacyl derivative, N-benzoylphenylalanyl derivative, N-acetylmethionine derivative, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm), N-2-picolylamino N′-oxide, N-1, 1-dimethylthiomethyleneamine, N-benzylideneamine, N-p-methoxybenzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine, N—(N′,N′-dimethylaminomethylene)amine, N,N′-isopropylidenediamine, N-p-nitrobenzylideneamine, N-salicylideneamine, N-5-chlorosalicylideneamine, N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine, N-borane derivative, N-diphenylborinic acid derivative, N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper chelate, N-zinc chelate, N-nitroamine, N-nitrosoamine, amine N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates, dibenzyl phosphoramidate, diphenyl phosphoramidate, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, and 3-nitropyridinesulfenamide (Npys).

In certain embodiments, the substituent present on an oxygen atom is an oxygen protecting group (also referred to as a hydroxyl protecting group). Oxygen protecting groups include, but are not limited to, —Raa, —N(Rbb)2, —C(═O)SRaa, —C(═O)Raa, —CO2Raa, —C(═O)N(Rbb)2, —C(═NRbb)Raa, —C(═NRbb)ORaa, —C(═NRbb)N(Rbb)2, —S(═O)Raa, —SO2Raa, Si(Raa)3, —P(Rcc)2, —P(Rcc)3, —P(═O)2Raa, —P(═O)(Raa)2, —P(═O)(ORcc)2, —P(═O)2N(Rbb)2, and —P(═O)(NRbb)2, wherein Raa, Rbb, and Rcc are as defined herein. Oxygen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999, incorporated herein by reference.

Exemplary oxygen protecting groups include, but are not limited to, methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl (CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxido, diphenylmethyl, p,p′-dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, α-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4′-bromophenacyloxyphenyl)diphenylmethyl, 4,4′,4″-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4′,4″-tris(levulinoyloxyphenyl)methyl, 4,4′,4″-tris(benzoyloxyphenyl)methyl, 3-(imidazol-1-yl)bis(4′,4″-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1′-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodisulfuran-2-yl, benzisothiazolyl S,S-dioxido, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (levulinate), 4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl 2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl) ethyl carbonate (Psec), 2-(triphenylphosphonio) ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate, 4-ethoxy-1-napththyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate, o-(methoxyacyl)benzoate, α-naphthoate, nitrate, alkyl N,N,N′,N′-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate, borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts).

In certain embodiments, the substituent present on an sulfur atom is an sulfur protecting group (also referred to as a thiol protecting group). Sulfur protecting groups include, but are not limited to, —Raa, —N(Rbb)2, —C(═O)SRaa, —C(═O)Raa, —CO2Raa, C(═O)N(Rbb)2, —C(═NRbb)Raa, —C(═NRbb)ORaa, —C(═NRbb)N(Rbb)2, —S(═O)Raa, —SO2Raa, —Si(Raa)3, —P(Rcc)2, —P(Rcc)3, —P(═O)2Raa, —P(═O)(Raa)2, —P(═O)(ORcc)2, —P(═O)2N(Rbb)2, and —P(═O)(NRbb)2, wherein Raa, Rbb, and Rcc are as defined herein. Sulfur protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999, incorporated herein by reference.

As used herein, a “leaving group” is an art-understood term referring to a molecular fragment that departs with a pair of electrons in heterolytic bond cleavage, wherein the molecular fragment is an anion or neutral molecule. See, for example, Smith, March's Advanced Organic Chemistry 6th ed. (501-502). Exemplary leaving groups include, but are not limited to, halo (e.g., chloro, bromo, iodo) and sulfonyl substituted hydroxyl groups (e.g., tosyl, mesyl, besyl).

Additional Definitions

Animal: As used herein, the term “animal” refers to any member of the animal kingdom. In some embodiments, “animal” refers to humans, at any stage of development. In some embodiments, “animal” refers to non-human animals, at any stage of development. In certain embodiments, the non-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, cattle, a primate, and/or a pig). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, insects, and/or worms. In some embodiments, an animal may be a transgenic animal, genetically-engineered animal, and/or a clone.

Approximately or about: As used herein, the term “approximately” or “about,” as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In certain embodiments, the term “approximately” or “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

Delivery: As used herein, the term “delivery” encompasses both local and systemic delivery. For example, delivery of mRNA encompasses situations in which an mRNA is delivered to a target tissue and the encoded protein is expressed and retained within the target tissue (also referred to as “local distribution” or “local delivery”), and situations in which an mRNA is delivered to a target tissue and the encoded protein is expressed and secreted into patient's circulation system (e.g., serum) and systematically distributed and taken up by other tissues (also referred to as “systemic distribution” or “systemic delivery).

Expression: As used herein, “expression” of a nucleic acid sequence refers to translation of an mRNA into a polypeptide, assemble multiple polypeptides (e.g., heavy chain or light chain of antibody) into an intact protein (e.g., antibody) and/or post-translational modification of a polypeptide or fully assembled protein (e.g., antibody). In this application, the terms “expression” and “production,” and grammatical equivalent, are used inter-changeably.

Improve, increase, or reduce: As used herein, the terms “improve,” “increase” or “reduce,” or grammatical equivalents, indicate values that are relative to a baseline measurement, such as a measurement in the same individual prior to initiation of the treatment described herein, or a measurement in a control subject (or multiple control subject) in the absence of the treatment described herein. A “control subject” is a subject afflicted with the same form of disease as the subject being treated, who is about the same age as the subject being treated.

In Vitro: As used herein, the term “in vitro” refers to events that occur in an artificial environment, e.g., in a test tube or reaction vessel, in cell culture, etc., rather than within a multi-cellular organism.

In Vivo: As used herein, the term “in vivo” refers to events that occur within a multi-cellular organism, such as a human and a non-human animal. In the context of cell-based systems, the term may be used to refer to events that occur within a living cell (as opposed to, for example, in vitro systems).

Isolated: As used herein, the term “isolated” refers to a substance and/or entity that has been (1) separated from at least some of the components with which it was associated when initially produced (whether in nature and/or in an experimental setting), and/or (2) produced, prepared, and/or manufactured by the hand of man. Isolated substances and/or entities may be separated from about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% of the other components with which they were initially associated. In some embodiments, isolated agents are about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% pure. As used herein, a substance is “pure” if it is substantially free of other components. As used herein, calculation of percent purity of isolated substances and/or entities should not include excipients (e.g., buffer, solvent, water, etc.).

Local distribution or delivery: As used herein, the terms “local distribution,” “local delivery,” or grammatical equivalent, refer to tissue specific delivery or distribution. Typically, local distribution or delivery requires a protein (e.g., enzyme) encoded by mRNAs be translated and expressed intracellularly or with limited secretion that avoids entering the patient's circulation system.

messenger RNA (mRNA): As used herein, the term “messenger RNA (mRNA)” refers to a polynucleotide that encodes at least one polypeptide. mRNA as used herein encompasses both modified and unmodified RNA. mRNA may contain one or more coding and non-coding regions. mRNA can be purified from natural sources, produced using recombinant expression systems and optionally purified, chemically synthesized, etc. Where appropriate, e.g., in the case of chemically synthesized molecules, mRNA can comprise nucleoside analogs such as analogs having chemically modified bases or sugars, backbone modifications, etc. An mRNA sequence is presented in the 5′ to 3′ direction unless otherwise indicated. In some embodiments, an mRNA is or comprises natural nucleosides (e.g., adenosine, guanosine, cytidine, uridine); nucleoside analogs (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-amino-adenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deaza-guanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, and 2-thiocytidine); chemically modified bases; biologically modified bases (e.g., methylated bases); intercalated bases; modified sugars (e.g., 2′-fluororibose, ribose, 2′-deoxyribose, arabinose, and hexose); and/or modified phosphate groups (e.g., phosphorothioates and 5′-N-phosphoramidite linkages).

Nucleic acid: As used herein, the term “nucleic acid,” in its broadest sense, refers to any compound and/or substance that is or can be incorporated into a polynucleotide chain. In some embodiments, a nucleic acid is a compound and/or substance that is or can be incorporated into a polynucleotide chain via a phosphodiester linkage. In some embodiments, “nucleic acid” refers to individual nucleic acid residues (e.g., nucleotides and/or nucleosides). In some embodiments, “nucleic acid” refers to a polynucleotide chain comprising individual nucleic acid residues. In some embodiments, “nucleic acid” encompasses RNA such as mRNA, siRNA, microRNA, as well as single and/or double-stranded DNA and/or cDNA.

Patient: As used herein, the term “patient” or “subject” refers to any organism to which a provided composition may be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and/or therapeutic purposes. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and/or humans). In some embodiments, a patient is a human. A human includes pre and post natal forms.

Pharmaceutically acceptable: The term “pharmaceutically acceptable” as used herein, refers to substances that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

Polymer: As used herein, a “polymer” refers to a compound comprised of at least 3 (e.g., at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, etc.) repeating covalently bound structural units.

Salt: As used herein, the term “salt” or “pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or rnalonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate. digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(C1-4alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium. quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, sulfonate and aryl sulfonate. Further pharmaceutically acceptable salts include salts formed from the quarternization of an amine using an appropriate electrophile, e.g., an alkyl halide, to form a quarternized alkylated amino salt.

Systemic distribution or delivery: As used herein, the terms “systemic distribution,” “systemic delivery,” or grammatical equivalent, refer to a delivery or distribution mechanism or approach that affect the entire body or an entire organism. Typically, systemic distribution or delivery is accomplished via body's circulation system, e.g., blood stream. Compared to the definition of “local distribution or delivery.”

Subject: As used herein, the term “subject” refers to a human or any non-human animal (e.g., mouse, rat, rabbit, dog, cat, cattle, swine, sheep, horse or primate). A human includes pre- and post-natal forms. In many embodiments, a subject is a human being. A subject can be a patient, which refers to a human presenting to a medical provider for diagnosis or treatment of a disease. The term “subject” is used herein interchangeably with “individual” or “patient.” A subject can be afflicted with or is susceptible to a disease or disorder but may or may not display symptoms of the disease or disorder.

Substantially: As used herein, the term “substantially” refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest. One of ordinary skill in the biological arts will understand that biological and chemical phenomena rarely, if ever, go to completion and/or proceed to completeness or achieve or avoid an absolute result. The term “substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

Target tissues: As used herein, the term “target tissues” refers to any tissue that is affected by a disease to be treated. In some embodiments, target tissues include those tissues that display disease-associated pathology, symptom, or feature.

DETAILED DESCRIPTION

The present invention provides, among other things, a novel class of biodegradable lipid compounds for improved in vivo delivery of therapeutic agents, such as nucleic acids. In particular, a biodegradable compound described herein may be used to as a cationic lipid, together with other non-cationic lipids, to formulate a lipid based nanoparticle (e.g., liposome) for encapsulating therapeutic agents, such as nucleic acids (e.g., DNA, siRNA, mRNA, microRNA) for therapeutic use.

Biodegradable Compounds

In some embodiments, a biodegradable compound according to the invention has a structure of formula I:

or a pharmaceutically acceptable salt thereof,

wherein:

-   -   each instance of X is independently O or S;     -   each instance of Y is independently O or S;     -   each instance of R¹ is independently hydrogen, optionally         substituted alkyl, optionally substituted alkenyl, optionally         substituted alkynyl, optionally substituted carbocyclyl,         optionally substituted heterocyclyl, optionally substituted         aryl, optionally substituted heteroaryl, halogen, —OR^(A1),         —N(R^(A1))₂, —SR^(A1), or a group of formula (iv):

-   -   L is an optionally substituted alkylene, optionally substituted         alkenylene, optionally substituted alkynylene, optionally         substituted heteroalkylene, optionally substituted         heteroalkenylene, optionally substituted heteroalkynylene,         optionally substituted carbocyclylene, optionally substituted         heterocyclylene, optionally substituted arylene, or optionally         substituted heteroarylene, or combination thereof, and each of         R⁶ and R⁷ is independently hydrogen, optionally substituted         alkyl, optionally substituted alkenyl, optionally substituted         alkynyl, optionally substituted carbocyclyl, optionally         substituted heterocyclyl, optionally substituted aryl,         optionally substituted heteroaryl, a nitrogen protecting group,         or a group of formula (i), (ii) or (iii); Formulae (i), (ii),         and (iii) are:

-   -   each instance of R′ is independently hydrogen or optionally         substituted alkyl;     -   X′ is O or S, or NR^(X);     -   R^(X) is hydrogen, optionally substituted alkyl, optionally         substituted alkenyl, optionally substituted alkynyl, optionally         substituted carbocyclyl, optionally substituted heterocyclyl,         optionally substituted aryl, optionally substituted heteroaryl,         or a nitrogen protecting group;     -   Y′ is O, S, or NR^(Y);     -   R^(Y) is hydrogen, optionally substituted alkyl, optionally         substituted alkenyl, optionally substituted alkynyl, optionally         substituted carbocyclyl, optionally substituted heterocyclyl,         optionally substituted aryl, optionally substituted heteroaryl,         or a nitrogen protecting group;     -   R^(P) is hydrogen, optionally substituted alkyl, optionally         substituted alkenyl, optionally substituted alkynyl, optionally         substituted carbocyclyl, optionally substituted heterocyclyl,         optionally substituted aryl, optionally substituted heteroaryl,         an oxygen protecting group when attached to an oxygen atom, a         sulfur protecting group when attached to a sulfur atom, or a         nitrogen protecting group when attached to a nitrogen atom;     -   R^(L) is optionally substituted C₁₋₅₀ alkyl, optionally         substituted C₂₋₅₀ alkenyl, optionally substituted C₂₋₅₀ alkynyl,         optionally substituted heteroC₁₋₅₀ alkyl, optionally substituted         heteroC₂₋₅₀ alkenyl, optionally substituted heteroC₂₋₅₀ alkynyl,         or a polymer;     -   and     -   each occurrence of R^(A1) is independently hydrogen, optionally         substituted alkyl, optionally substituted alkenyl, optionally         substituted alkynyl, optionally substituted carbocyclyl,         optionally substituted heterocyclyl, optionally substituted         aryl, optionally substituted heteroaryl, an oxygen protecting         group when attached to an oxygen atom, a sulfur protecting group         when attached to an sulfur atom, a nitrogen protecting group         when attached to a nitrogen atom, or two R^(A1) groups, together         with the nitrogen atom to which they are attached, are joined to         form an optionally substituted heterocyclic or optionally         substituted heteroaryl ring.

In certain embodiments, a group of formula (i) represents a group of formula (i-a) or a group of formula (i-b):

wherein each variable is independently as defined above and described herein. In some embodiments, a group of formula (i) is a group of formula (i-a). In some embodiments, a group of formula (i) is a group of formula (i-b).

In some embodiments, at least one instance of R¹ is a group of formula (iv). In some embodiments, at least one instance of R¹ is a group of formula (iv), wherein at least one of R⁶ and R⁷ is a group of formula (i), (ii) or (iii). In some embodiments, at least one instance of R¹ is a group of formula (iv), wherein each of R⁶ and R⁷ is independently a group of formula (i), (ii) or (iii).

In some embodiments, each R¹ is independently a group of formula (iv). In some embodiments, each R¹ is independently a group of formula (iv), wherein at least one of R⁶ and R⁷ is a group of formula (i), (ii) or (iii). In some embodiments, each R¹ is independently a group of formula (iv), wherein each of R⁶ and R⁷ is independently a group of formula (i), (ii) or (iii). In some embodiments, each R¹ is independently a group of formula (iv), wherein each of R⁶ and R⁷ is independently a group of formula (i). In some embodiments, each R¹ is independently a group of formula (iv), wherein each of R⁶ and R⁷ is independently a group of formula (ii). In some embodiments, each R¹ is independently a group of formula (iv), wherein each of R⁶ and R⁷ is independently a group of formula (iii). In some embodiments, each R¹ is independently a group of formula (iv), wherein each of R⁶ and R⁷ is independently a group of formula (i-a). In some embodiments, each R¹ is independently a group of formula (iv), wherein each of R⁶ and R⁷ is independently a group of formula (i-b).

In some embodiments, each instance of R′ is hydrogen.

In some embodiments, L is an optionally substituted alkylene.

In some embodiments, a group of formula (iv) is of formula

wherein q is an integer between 1 and 50, inclusive, and each of R⁶ and R⁷ is independently as defined above and described herein.

In certain embodiments, at least one instance of Q is O. In certain embodiments, each instance of Q is O. In certain embodiments, at least one instance of Q is S. In certain embodiments, each instance of Q is S.

In some embodiments, R^(Q) is hydrogen. In some embodiments, R^(Q) is optionally substituted alkyl. In some embodiments, R^(Q) is optionally substituted alkenyl. In some embodiments, R^(Q) is optionally substituted alkynyl. In some embodiments, R^(Q) is carbocyclyl. In some embodiments, R^(Q) is optionally substituted heterocyclyl. In some embodiments, R^(Q) is optionally substituted aryl. In some embodiments, R^(Q) is optionally substituted heteroaryl. In some embodiments, R^(Q) is a nitrogen protecting group. In some embodiments, R^(Q) is a group of formula (i), (ii) or (iii). In some embodiments, R^(Q) is a group of formula (i). In some embodiments, R^(Q) is a group of formula (ii). In some embodiments, R^(Q) is a group of formula (iii).

As generally defined above, each instance of R¹ is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, halogen, —OR^(A1), —N(R^(A1))₂, or —SR^(A1), or a group of formula (iv), wherein each of R^(A1) and formula (iv) is independently as defined above and described herein.

In some embodiments, one R¹ is not hydrogen. In some embodiments, both of R¹ are not hydrogen.

In certain embodiments, R¹ is optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl. In certain embodiments, at least one instance of R¹ is optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl.

In certain embodiments, R¹ is optionally substituted alkyl; e.g., optionally substituted C₁₋₆alkyl, optionally substituted C₂₋₆alkyl, optionally substituted C₃₋₆alkyl, optionally substituted C₄₋₆alkyl, optionally substituted C₄₋₅alkyl, or optionally substituted C₃₋₄alkyl. In certain embodiments, at least one instance of R¹ is optionally substituted alkyl; e.g., optionally substituted C₁₋₆alkyl, optionally substituted C₂₋₆alkyl, optionally substituted C₃₋₆alkyl, optionally substituted C₄₋₆alkyl, optionally substituted C₄₋₅alkyl, or optionally substituted C₃₋₄alkyl.

In certain embodiments, R¹ is optionally substituted alkenyl, e.g., optionally substituted C₂₋₆alkenyl, optionally substituted C₃₋₆alkenyl, optionally substituted C₄₋₆alkenyl, optionally substituted C₄₋₅alkenyl, or optionally substituted C₃₋₄alkenyl. In certain embodiments, at least one instance of R¹ is optionally substituted alkenyl, e.g., optionally substituted C₂₋₆alkenyl, optionally substituted C₃₋₆alkenyl, optionally substituted C₄₋₆alkenyl, optionally substituted C₄₋₅alkenyl, or optionally substituted C₃₋₄alkenyl.

In certain embodiments, R¹ is optionally substituted alkynyl, e.g., optionally substituted C₂₋₆alkynyl, optionally substituted C₃₋₆alkynyl, optionally substituted C₄₋₆alkynyl, optionally substituted C₄₋₅alkynyl, or optionally substituted C₃₋₄alkynyl. In certain embodiments, at least one instance of R¹ is optionally substituted alkynyl, e.g., optionally substituted C₂₋₆alkynyl, optionally substituted C₃₋₆alkynyl, optionally substituted C₄₋₆alkynyl, optionally substituted C₄₋₅alkynyl, or optionally substituted C₃₋₄alkynyl.

In certain embodiments, R¹ is optionally substituted carbocyclyl, e.g., optionally substituted C₃₋₁₀ carbocyclyl, optionally substituted C₅₋₈ carbocyclyl, optionally substituted C₅₋₆ carbocyclyl, optionally substituted C₅ carbocyclyl, or optionally substituted C₆ carbocyclyl. In certain embodiments, at least one instance of R¹ is optionally substituted carbocyclyl, e.g., optionally substituted C₃₋₁₀ carbocyclyl, optionally substituted C₅₋₈ carbocyclyl, optionally substituted C₅₋₆ carbocyclyl, optionally substituted C₅ carbocyclyl, or optionally substituted C₆ carbocyclyl.

In some embodiments, R¹ is optionally substituted heterocyclyl, e.g., optionally substituted 3-14 membered heterocyclyl, optionally substituted 3-10 membered heterocyclyl, optionally substituted 5-8 membered heterocyclyl, optionally substituted 5-6 membered heterocyclyl, optionally substituted 5-membered heterocyclyl, or optionally substituted 6-membered heterocyclyl. In certain embodiments, at least one instance of R¹ is optionally substituted heterocyclyl, e.g., optionally substituted 3-14 membered heterocyclyl, optionally substituted 3-10 membered heterocyclyl, optionally substituted 5-8 membered heterocyclyl, optionally substituted 5-6 membered heterocyclyl, optionally substituted 5-membered heterocyclyl, or optionally substituted 6-membered heterocyclyl.

In some embodiments, R¹ is optionally substituted aryl. In some embodiments, R¹ is optionally substituted phenyl. In some embodiments, R¹ is phenyl. In some embodiments, R¹ is substituted phenyl. In certain embodiments, at least one instance of R¹ is optionally substituted aryl, e.g., optionally substituted phenyl.

In some embodiments, R¹ is optionally substituted heteroaryl, e.g., optionally substituted 5-14 membered heteroaryl, optionally substituted 5-10 membered heteroaryl, optionally substituted 5-6 membered heteroaryl, optionally substituted 5 membered heteroaryl, or optionally substituted 6 membered heteroaryl. In certain embodiments, at least one instance of R¹ is optionally substituted heteroaryl, e.g., optionally substituted 5-14 membered heteroaryl, optionally substituted 5-10 membered heteroaryl, optionally substituted 5-6 membered heteroaryl, optionally substituted 5 membered heteroaryl, or optionally substituted 6 membered heteroaryl.

In some embodiments, R¹ is halogen. In some embodiments, R¹ is —F. In some embodiments, R¹ is —Cl. In some embodiments, R¹ is —Br. In some embodiments, R¹ is —I.

In some embodiments, R¹ is —OR^(A1), wherein R^(A1) is as defined above and described herein. In some embodiments, R¹ is —N(R^(A1))₂, wherein each R^(A1) is independently as defined above and described herein. In some embodiments, R¹ is —SR^(A1), wherein R^(A1) is as defined above and described herein.

In some embodiments, an R¹ alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl group may be substituted. In some embodiments, an R¹ alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl group may be substituted with an optionally substituted amino group. In some embodiments, an R¹ alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl group may be substituted with an optionally substituted hydroxyl group. In some embodiments, an R¹ alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl group may be substituted with an optionally substituted thiol group. In any of the above embodiments, an R¹ alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl group may be substituted, for example, with an optionally substituted amino group (e.g., —NR⁶R⁷), an optionally substituted hydroxyl group (e.g., —OR⁶), an optionally substituted thiol group (e.g., —SR⁶), or with a group of formula (i), (ii), or (iii), wherein each instance of R⁶ and R⁷ is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group when attached to a nitrogen atom, an oxygen protecting group when attached to an oxygen atom, and a sulfur protecting group when attached to a sulfur atom, or a group of formula (i), (ii), or (iii).

In some embodiments, R¹ is an optionally substituted natural amino acid side chain. In some embodiments, R¹ is a natural amino acid side chain. In some embodiments, R¹ is an optionally substituted unnatural amino acid side chain. In some embodiments, R¹ is an unnatural amino acid side chain.

In certain embodiments, each instance of R¹ is the same. In certain embodiments, at least one R¹ group is different. In certain embodiments, each R¹ group is different.

In certain embodiments, R¹ is an alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl group substituted with an amino group of the formula —NR⁶R⁷.

In certain embodiments, R¹ is a group of formula (iv):

wherein: L is an optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted heteroalkenylene, optionally substituted heteroalkynylene, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene, or combination thereof; and each of R⁶ and R⁷ is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group, or a group of formula (i), (ii) or (iii):

wherein each of R′, Y′, R^(P), R^(L) and X′ is independently as defined above and described herein.

In some embodiments, at least one instance of R¹ is an alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl group substituted with an amino group of the formula —NR⁶R⁷. In some embodiments, at least one instance of R¹ is a group of formula (iv). In some embodiments, at least one instance of R¹ is a group of formula (iv), wherein at least one instance of R⁶ and R⁷ is a group of the formula (i), (ii) or (iii). In some embodiments, at least one instance of R¹ is a group of formula (iv), wherein each instance of R⁶ and R⁷ is a group of the formula (i), (ii) or (iii). In some embodiments, at least one instance of R¹ is a group of formula (iv), wherein each instance of R⁶ and R⁷ is a group of the formula (i). In some embodiments, at least one instance of R¹ is a group of formula (iv), wherein each instance of R⁶ and R⁷ is a group of the formula (ii). In some embodiments, at least one instance of R¹ is a group of formula (iv), wherein each instance of R⁶ and R⁷ is a group of the formula (iii).

In some embodiments, each instance of R¹ is a group of formula (iv). In some embodiments, each instance of R¹ is a group of formula (iv), wherein each instance of R⁶ and R⁷ is a group of the formula (i), (ii) or (iii). In some embodiments, each instance of R¹ is a group of formula (iv), wherein each instance of R⁶ and R⁷ is a group of the formula (i), (ii) or (iii). In some embodiments, each instance of R¹ is a group of formula (iv), wherein each instance of R⁶ and R⁷ is a group of the formula (i). In some embodiments, each instance of R¹ is a group of formula (iv), wherein each instance of R⁶ and R⁷ is a group of the formula (ii). In some embodiments, each instance of R¹ is a group of formula (iv), wherein each instance of R⁶ and R⁷ is a group of the formula (iii).

In certain embodiments, at least two instances of R¹ is a group of formula (iv). In certain embodiments, at least three instances of R¹ is a group of formula (iv). In certain embodiments, at least four instances of R¹ is a group of formula (iv). In certain embodiments, at least five instances of R¹ is a group of formula (iv). In certain embodiments, at least six instances of R¹ is a group of formula (iv). In certain embodiments, at least seven instances of R¹ is a group of formula (iv). In certain embodiments, at least eight instances of R¹ is a group of formula (iv). In certain embodiments, at least nine instances of R¹ is a group of formula (iv). In certain embodiments, each instance of R¹ is a group of formula (iv).

In certain embodiments, L is an optionally substituted alkylene; e.g., optionally substituted C₁₋₅₀alkylene, optionally substituted C₁₋₄₀alkylene, optionally substituted C₁₋₃₀alkylene, optionally substituted C₁₋₂₀alkylene, optionally substituted C₄₋₂₀alkylene, optionally substituted C₆₋₂₀alkylene, optionally substituted C₈₋₂₀alkylene, optionally substituted C₁₀₋₂₀alkylene, optionally substituted C₁₋₆alkylene, optionally substituted C₂₋₆alkylene, optionally substituted C₃₋₆alkylene, optionally substituted C₄₋₆alkylene, optionally substituted C₄₋₅alkylene, or optionally substituted C₃₋₄alkylene. In some embodiments, L is optionally substituted C₁ alkylene. In some embodiments, L is optionally substituted C₂ alkylene. In some embodiments, L is optionally substituted C₃ alkylene. In some embodiments, L is optionally substituted C₄ alkylene. In some embodiments, L is optionally substituted C₅ alkylene. In some embodiments, L is optionally substituted C₆ alkylene. In some embodiments, L is optionally substituted C₇ alkylene. In some embodiments, L is optionally substituted C₈ alkylene. In some embodiments, L is optionally substituted C₉ alkylene. In some embodiments, L is optionally substituted C₁₀ alkylene. In some embodiments, L is —CH₂—. In some embodiments, L is —(CH₂)₂—. In some embodiments, L is —(CH₂)₃—. In some embodiments, L is —(CH₂)₄—. In some embodiments, L is —(CH₂)₅—. In some embodiments, L is —(CH₂)₆—. In some embodiments, L is —(CH₂)₇—. In some embodiments, L is —(CH₂)₈—. In some embodiments, L is —(CH₂)₉—. In some embodiments, L is —(CH₂)₁₀—.

In certain embodiments, L is an optionally substituted alkenylene, e.g., optionally substituted C₂₋₅₀alkenylene, optionally substituted C₂₋₄₀alkenylene, optionally substituted C₂₋₃₀alkenylene, optionally substituted C₂₋₂₀alkenylene, optionally substituted C₄₋₂₀alkenylene, optionally substituted C₆₋₂₀alkenylene, optionally substituted C₈₋₂₀alkenylene, optionally substituted C₁₀₋₂₀alkenylene, optionally substituted C₂₋₆alkenylene, optionally substituted C₃₋₆alkenylene, optionally substituted C₄₋₆alkenylene, optionally substituted C₄₋₅alkenylene, or optionally substituted C₃₋₄alkenylene.

In certain embodiments, L is an optionally substituted alkynylene, e.g., optionally substituted C₂₋₅₀alkynylene, optionally substituted C₂₋₄₀alkynylene, optionally substituted C₂₋₃₀alkynylene, optionally substituted C₂₋₂₀alkynylene, optionally substituted C₄₋₂₀alkynylene, optionally substituted C₆₋₂₀alkynylene, optionally substituted C₈₋₂₀alkynylene, optionally substituted C₁₀₋₂₀alkynylene, optionally substituted C₂₋₆alkynylene, optionally substituted C₃₋₆alkynylene, optionally substituted C₄₋₆alkynylene, optionally substituted C₄₋₅alkynylene, or optionally substituted C₃₋₄alkynylene.

In certain embodiments, L is an optionally substituted heteroalkylene; e.g., optionally substituted heteroC₁₋₅₀alkylene, optionally substituted heteroC₁₋₄₀alkylene, optionally substituted heteroC₁₋₃₀alkylene, optionally substituted heteroC₁₋₂₀alkylene, optionally substituted heteroC₄₋₂₀alkylene, optionally substituted heteroC₆₋₂₀alkylene, optionally substituted heteroC₈₋₂₀alkylene, optionally substituted heteroC₁₀₋₂₀alkylene, optionally substituted heteroC₁₋₆alkylene, optionally substituted heteroC₂₋₆alkylene, optionally substituted heteroC₃₋₆alkylene, optionally substituted heteroC₄₋₆alkylene, optionally substituted heteroC₄₋₅alkylene, or optionally substituted heteroC₃₋₄alkylene. In some embodiments, L is optionally substituted heteroC₂alkylene. In some embodiments, L is optionally substituted heteroC₃alkylene. In some embodiments, L is optionally substituted heteroC₄alkylene. In some embodiments, L is optionally substituted heteroC₅alkylene. In some embodiments, L is optionally substituted heteroC₆alkylene. In some embodiments, L is optionally substituted heteroC₇alkylene. In some embodiments, L is optionally substituted heteroC₅alkylene. In some embodiments, L is optionally substituted heteroC₉alkylene. In some embodiments, L is optionally substituted heteroC₁₀alkylene.

In certain embodiments, L is an optionally substituted heteroalkenylene, e.g., optionally substituted heteroC₂₋₅₀alkenylene, optionally substituted heteroC₂₋₄₀alkenylene, optionally substituted heteroC₂₋₃₀alkenylene, optionally substituted heteroC₂₋₂₀alkenylene, optionally substituted heteroC₄₋₂₀alkenylene, optionally substituted heteroC₆₋₂₀alkenylene, optionally substituted heteroC₅₋₂₀alkenylene, optionally substituted heteroC₁₀₋₂₀alkenylene, optionally substituted heteroC₂₋₆alkenylene, optionally substituted heteroC₃₋₆alkenylene, optionally substituted heteroC₄₋₆alkenylene, optionally substituted heteroC₄₋₅alkenylene, or optionally substituted heteroC₃₋₄alkenylene.

In certain embodiments, L is an optionally substituted heteroalkynylene, e.g., optionally substituted heteroC₂₋₅₀alkynylene, optionally substituted heteroC₂₋₄₀alkynylene, optionally substituted heteroC₂₋₃₀alkynylene, optionally substituted heteroC₂₋₂₀alkynylene, optionally substituted heteroC₄₋₂₀alkynylene, optionally substituted heteroC₆₋₂₀alkynylene, optionally substituted heteroC₈₋₂₀alkynylene, optionally substituted heteroC₁₀₋₂₀alkynylene, optionally substituted heteroC₂₋₆alkynylene, optionally substituted heteroC₃₋₆alkynylene, optionally substituted heteroC₄₋₆alkynylene, optionally substituted heteroC₄₋₅alkynylene, or optionally substituted heteroC₃₋₄alkynylene.

In certain embodiments, L is an optionally substituted carbocyclylene, e.g., optionally substituted C₃₋₁₀carbocyclylene, optionally substituted C₅₋₈carbocyclylene, optionally substituted C₅₋₆carbocyclylene, optionally substituted C₅carbocyclylene, or optionally substituted C₆carbocyclylene.

In certain embodiments, L is an optionally substituted heterocyclylene, e.g., optionally substituted 3-14 membered heterocyclylene, optionally substituted 3-10 membered heterocyclylene, optionally substituted 5-8 membered heterocyclylene, optionally substituted 5-6 membered heterocyclylene, optionally substituted 5-membered heterocyclylene, or optionally substituted 6-membered heterocyclylene.

In certain embodiments, L is an optionally substituted arylene, e.g., optionally substituted phenylene. In some embodiments, L is optionally substituted phenylene. In some embodiments, L is substituted phenylene. In some embodiments, L is unsubstituted phenylene.

In certain embodiments, L is an optionally substituted heteroarylene, e.g., optionally substituted 5-14 membered heteroarylene, optionally substituted 5-10 membered heteroarylene, optionally substituted 5-6 membered heteroarylene, optionally substituted 5-membered heteroarylene, or optionally substituted 6-membered heteroarylene.

In certain embodiments, wherein L is an optionally substituted alkylene group, the group of formula (iv) is a group of the formula

wherein q is an integer between 1 and 50, inclusive, and each of R⁶ and R⁷ is independently as defined above and described herein.

In certain embodiments, q is an integer between 1 and 40, inclusive. In certain embodiments, q is an integer between 1 and 30, inclusive. In certain embodiments, q is an integer between 1 and 20, inclusive. In certain embodiments, q is an integer between 1 and 10, inclusive. In certain embodiments, q is an integer between 4 and 20, inclusive. In certain embodiments, q is an integer between 6 and 20, inclusive. In certain embodiments, q is an integer between 2 and 10, inclusive. In certain embodiments, q is an integer between 2 and 9, inclusive. In certain embodiments, q is an integer between 2 and 8, inclusive. In certain embodiments, q is an integer between 2 and 7, inclusive. In certain embodiments, q is an integer between 2 and 6, inclusive. In certain embodiments, q is an integer between 2 and 5, inclusive. In certain embodiments, q is an integer between 2 and 4, inclusive. In certain embodiments, q is an integer between 3 and 10, inclusive. In certain embodiments, q is an integer between 3 and 8, inclusive. In certain embodiments, q is an integer between 3 and 7, inclusive. In certain embodiments, q is an integer between 3 and 6, inclusive. In certain embodiments, q is an integer between 3 and 5, inclusive. In certain embodiments, q is 3 or 4. In certain embodiments, q is an integer between 3 and 9, inclusive. In certain embodiments, q is an integer between 8 and 20, inclusive. In certain embodiments, q is 1. In certain embodiments, q is 2. In certain embodiments, q is 3. In certain embodiments, q is 4. In certain embodiments, q is 5. In certain embodiments, q is 6. In certain embodiments, q is 7. In certain embodiments, q is 8. In certain embodiments, q is 9. In certain embodiments, q is 10.

As generally defined above, each R⁶ is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group, or a group of formula (i), (ii) or (iii).

In some embodiments, R⁶ is hydrogen.

In some embodiments, R⁶ is optionally substituted alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₅₀ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₄₀ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₃₀ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₂₀ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₉ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₈ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₇ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₆ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₅ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₄ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₃ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₂ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₁ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₀ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₉ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₈ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₇ alkyl. In some embodiments, R⁶ is optionally substituted C₂₋₆ alkyl.

In some embodiments, R⁶ is optionally substituted C₄₋₅₀ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₄₀ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₃₀ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₂₀ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₉ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₈ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₇ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₆ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₅ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₄ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₃ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₂ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₁ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₀ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₉ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₈ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₇ alkyl. In some embodiments, R⁶ is optionally substituted C₄₋₆ alkyl.

In some embodiments, R⁶ is optionally substituted C₆₋₅₀ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₄₀ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₃₀ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₂₀ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₉ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₈ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₇ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₆ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₅ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₄ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₃ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₂ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₁ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₀ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₉ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₈ alkyl. In some embodiments, R⁶ is optionally substituted C₆₋₇ alkyl.

In some embodiments, R⁶ is optionally substituted C₈₋₅₀ alkyl. In some embodiments, R⁶ is optionally substituted C₈₋₄₀ alkyl. In some embodiments, R⁶ is optionally substituted C₈₋₃₀ alkyl. In some embodiments, R⁶ is optionally substituted C₈₋₂₀ alkyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₉ alkyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₈ alkyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₇ alkyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₆ alkyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₅ alkyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₄ alkyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₃ alkyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₂ alkyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₁ alkyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₀ alkyl. In some embodiments, R⁶ is optionally substituted C₈₋₉ alkyl.

In some embodiments, R⁶ is optionally substituted C₉₋₅₀ alkyl. In some embodiments, R⁶ is optionally substituted C₉₋₄₀ alkyl. In some embodiments, R⁶ is optionally substituted C₉₋₃₀ alkyl. In some embodiments, R⁶ is optionally substituted C₉₋₂₀ alkyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₉ alkyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₈ alkyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₇ alkyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₆ alkyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₅ alkyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₄ alkyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₃ alkyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₂ alkyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₁ alkyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₀ alkyl.

In some embodiments, R⁶ is optionally substituted C₁₀₋₅₀ alkyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₄₀ alkyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₃₀ alkyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₂₀ alkyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₉ alkyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₈alkyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₇ alkyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₆ alkyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₅ alkyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₄ alkyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₃ alkyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₂ alkyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₁ alkyl.

In some embodiments, R⁶ is optionally substituted C₁₁₋₅₀ alkyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₄₀ alkyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₃₀ alkyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₂₀ alkyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₉ alkyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₈ alkyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₇ alkyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₆ alkyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₅ alkyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₄ alkyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₃ alkyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₂ alkyl.

In some embodiments, R⁶ is optionally substituted C₁₂₋₅₀ alkyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₄₀ alkyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₃₀ alkyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₂₀ alkyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₉ alkyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₈ alkyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₇ alkyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₆ alkyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₅ alkyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₄ alkyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₃ alkyl.

In some embodiments, R⁶ is optionally substituted C₆ alkyl. In some embodiments, R⁶ is optionally substituted C₇ alkyl. In some embodiments, R⁶ is optionally substituted C₈ alkyl. In some embodiments, R⁶ is optionally substituted C₉ alkyl. In some embodiments, R⁶ is optionally substituted C₁₀alkyl. In some embodiments, R⁶ is optionally substituted C₁₁ alkyl. In some embodiments, R⁶ is optionally substituted C₁₂ alkyl. In some embodiments, R⁶ is optionally substituted C₁₃ alkyl. In some embodiments, R⁶ is optionally substituted C₁₄ alkyl. In some embodiments, R⁶ is optionally substituted C₁₅ alkyl. In some embodiments, R⁶ is optionally substituted C₁₆ alkyl. In some embodiments, R⁶ is optionally substituted C₁₇ alkyl. In some embodiments, R⁶ is optionally substituted C₁₈ alkyl. In some embodiments, R⁶ is optionally substituted C₁₉ alkyl. In some embodiments, R⁶ is optionally substituted C₂₀ alkyl.

In some embodiments, for example, in any of the above embodiments, R⁶ is a substituted alkyl group. In some embodiments, R⁶ is an unsubstituted alkyl group. In some embodiments, R⁶ is an optionally substituted straight-chain alkyl group. In some embodiments, R⁶ is a substituted straight-chain alkyl group. In some embodiments, R⁶ is an unsubstituted straight-chain alkyl group. In some embodiments, R⁶ is an optionally substituted branched alkyl group. In some embodiments, R⁶ is a substituted branched alkyl group. In some embodiments, R⁶ is an unsubstituted branched alkyl group.

In some embodiments, R⁶ is optionally substituted alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₅₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₄₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₃₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₂₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₉ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₈ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₇ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₆ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₅ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₄ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₃ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₂ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₁ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₉ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₈ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₇ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₋₆ alkenyl.

In some embodiments, R⁶ is optionally substituted C₄₋₅₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₄₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₃₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₂₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₉ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₈ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₇ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₆ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₅ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₄ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₃ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₂ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₁ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₉ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₈ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₇ alkenyl. In some embodiments, R⁶ is optionally substituted C₄₋₆ alkenyl.

In some embodiments, R⁶ is optionally substituted C₆₋₅₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₄₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₃₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₂₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₉ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₈ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₇ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₆ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₅ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₄ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₃ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₂ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₁ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₉ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₈ alkenyl. In some embodiments, R⁶ is optionally substituted C₆₋₇ alkenyl.

In some embodiments, R⁶ is optionally substituted C₈₋₅₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₈₋₄₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₈₋₃₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₈₋₂₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₉ alkenyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₈ alkenyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₇ alkenyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₆ alkenyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₅ alkenyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₄ alkenyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₃ alkenyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₂ alkenyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₁ alkenyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₈₋₉ alkenyl.

In some embodiments, R⁶ is optionally substituted C₉₋₅₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₉₋₄₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₉₋₃₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₉₋₂₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₉ alkenyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₈ alkenyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₇ alkenyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₆ alkenyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₅ alkenyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₄ alkenyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₃ alkenyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₂ alkenyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₁ alkenyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₀ alkenyl.

In some embodiments, R⁶ is optionally substituted C₁₀₋₅₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₄₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₃₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₂₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₉ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₈ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₇ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₆ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₅ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₄ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₃ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₂ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₁ alkenyl.

In some embodiments, R⁶ is optionally substituted C₁₁₋₅₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₄₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₃₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₂₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₉ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₁ s alkenyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₇ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₆ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₅ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₄ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₃ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₂ alkenyl.

In some embodiments, R⁶ is optionally substituted C₁₂₋₅₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₄₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₃₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₂₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₉ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₈ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₇ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₆ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₅ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₄ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₃ alkenyl.

In some embodiments, R⁶ is optionally substituted C₆ alkenyl. In some embodiments, R⁶ is optionally substituted C₇ alkenyl. In some embodiments, R⁶ is optionally substituted C₈ alkenyl. In some embodiments, R⁶ is optionally substituted C₉ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₀ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₁ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₂ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₃ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₄ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₅ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₆ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₇ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₈ alkenyl. In some embodiments, R⁶ is optionally substituted C₁₉ alkenyl. In some embodiments, R⁶ is optionally substituted C₂₀ alkenyl.

In some embodiments, for example, in any of the above embodiments, R⁶ is a substituted alkenyl group. In some embodiments, R⁶ is an unsubstituted alkenyl group. In some embodiments, R⁶ is an optionally substituted straight-chain alkenyl group. In some embodiments, R⁶ is a substituted straight-chain alkenyl group. In some embodiments, R⁶ is an unsubstituted straight-chain alkenyl group. In some embodiments, R⁶ is an optionally substituted branched alkenyl group. In some embodiments, R⁶ is a substituted branched alkenyl group. In some embodiments, R⁶ is an unsubstituted branched alkenyl group.

In some embodiments, R⁶ is optionally substituted alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₅₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₄₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₃₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₂₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₉ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₈ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₇ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₆ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₅ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₄ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₃ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₂ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₁ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₁₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₉ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₈ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₇ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₋₆ alkynyl.

In some embodiments, R⁶ is optionally substituted C₄₋₅₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₄₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₃₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₂₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₉ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₈ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₇ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₆ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₅ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₄ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₃ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₂ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₁ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₁₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₉ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₈ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₇ alkynyl. In some embodiments, R⁶ is optionally substituted C₄₋₆ alkynyl.

In some embodiments, R⁶ is optionally substituted C₆₋₅₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₄₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₃₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₂₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₉ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₈ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₇ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₆ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₅ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₄ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₃ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₂ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₁ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₁₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₉ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₈ alkynyl. In some embodiments, R⁶ is optionally substituted C₆₋₇ alkynyl.

In some embodiments, R⁶ is optionally substituted C₈₋₅₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₈₋₄₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₈₋₃₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₈₋₂₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₉ alkynyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₈ alkynyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₇ alkynyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₆ alkynyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₅ alkynyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₄ alkynyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₃ alkynyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₂ alkynyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₁ alkynyl. In some embodiments, R⁶ is optionally substituted C₈₋₁₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₈₋₉ alkynyl.

In some embodiments, R⁶ is optionally substituted C₉₋₅₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₉₋₄₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₉₋₃₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₉₋₂₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₉ alkynyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₈ alkynyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₇ alkynyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₆ alkynyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₅ alkynyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₄ alkynyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₃ alkynyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₂ alkynyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₁ alkynyl. In some embodiments, R⁶ is optionally substituted C₉₋₁₀ alkynyl.

In some embodiments, R⁶ is optionally substituted C₁₀₋₅₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₄₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₃₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₂₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₉ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₈ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₇ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₆ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₅ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₄ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₃ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₂ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₀₋₁₁ alkynyl.

In some embodiments, R⁶ is optionally substituted C₁₁₋₅₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₄₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₃₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₋₂₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₉ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₁ s alkynyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₇ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₆ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₅ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₄ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₃ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₁₋₁₂ alkynyl.

In some embodiments, R⁶ is optionally substituted C₁₂₋₅₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₄₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₃₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₂₀ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₉ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₈ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₇ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₆ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₅ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₄ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₂₋₁₃ alkynyl.

In some embodiments, R⁶ is optionally substituted C₆ alkynyl. In some embodiments, R⁶ is optionally substituted C₇ alkynyl. In some embodiments, R⁶ is optionally substituted C₈ alkynyl. In some embodiments, R⁶ is optionally substituted C₉ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₀alkynyl. In some embodiments, R⁶ is optionally substituted C₁₁ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₂ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₃ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₄ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₅ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₆ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₇ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₈ alkynyl. In some embodiments, R⁶ is optionally substituted C₁₉ alkynyl. In some embodiments, R⁶ is optionally substituted C₂₀ alkynyl.

In some embodiments, for example, in any of the above embodiments, R⁶ is a substituted alkynyl group. In some embodiments, R⁶ is an unsubstituted alknyl group. In some embodiments, R⁶ is an optionally substituted straight-chain alkynyl group. In some embodiments, R⁶ is a substituted straight-chain alkynyl group. In some embodiments, R⁶ is an unsubstituted straight-chain alkynyl group. In some embodiments, R⁶ is an optionally substituted branched alkynyl group. In some embodiments, R⁶ is a substituted branched alkynyl group. In some embodiments, R⁶ is an unsubstituted branched alkynyl group.

In some embodiments, R⁶ is optionally substituted carbocyclyl. In some embodiments, R⁶ is optionally substituted heterocyclyl. In some embodiments, R⁶ is optionally substituted aryl. In some embodiments, R⁶ is optionally substituted heteroaryl. In some embodiments, R⁶ is a nitrogen protecting group.

In some embodiments, R⁶ is a group of formula (i). In some embodiments, R⁶ is a group of formula (i-a). In some embodiments, R⁶ is a group of formula

In some embodiments, R⁶ is a group of formula (i-b). In some embodiments, R⁶ is a group of formula (ii). In some embodiments, R⁶ is a group of formula (iii).

In some embodiments, R⁶ is substituted with one or more hydroxyl groups. In some embodiments, R⁶ is substituted with one hydroxyl group. In some embodiments, R⁶ is substituted with one 2-hydroxyl group (C1 is the carbon atom directly bonded to the nitrogen atom depicted in formula (iv)).

As generally defined above, each R⁷ is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group, or a group of formula (i), (ii) or (iii).

In some embodiments, R⁷ is hydrogen.

In some embodiments, R⁷ is optionally substituted alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₅₀ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₄₀ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₃₀ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₂₀ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₉ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₈ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₇ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₆ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₅ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₄ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₃ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₂ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₁ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₀ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₉ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₈ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₇ alkyl. In some embodiments, R⁷ is optionally substituted C₂₋₆ alkyl.

In some embodiments, R⁷ is optionally substituted C₄₋₅₀ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₄₀ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₃₀ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₂₀ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₉ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₈ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₇ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₆ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₅ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₄ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₃ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₂ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₁ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₀ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₉ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₈ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₇ alkyl. In some embodiments, R⁷ is optionally substituted C₄₋₆ alkyl.

In some embodiments, R⁷ is optionally substituted C₆₋₅₀ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₄₀ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₃₀ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₂₀ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₉ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₈ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₇ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₆ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₅ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₄ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₃ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₂ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₁ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₀ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₉ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₈ alkyl. In some embodiments, R⁷ is optionally substituted C₆₋₇ alkyl.

In some embodiments, R⁷ is optionally substituted C₅₋₅₀ alkyl. In some embodiments, R⁷ is optionally substituted C₈₋₄₀ alkyl. In some embodiments, R⁷ is optionally substituted C₈₋₃₀ alkyl. In some embodiments, R⁷ is optionally substituted C₈₋₂₀ alkyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₉ alkyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₈ alkyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₇ alkyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₆ alkyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₅ alkyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₄ alkyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₃ alkyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₂ alkyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₁ alkyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₀ alkyl. In some embodiments, R⁷ is optionally substituted C₈₋₉ alkyl.

In some embodiments, R⁷ is optionally substituted C₉₋₅₀ alkyl. In some embodiments, R⁷ is optionally substituted C₉₋₄₀ alkyl. In some embodiments, R⁷ is optionally substituted C₉₋₃₀ alkyl. In some embodiments, R⁷ is optionally substituted C₉₋₂₀ alkyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₉ alkyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₈ alkyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₇ alkyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₆ alkyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₅ alkyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₄ alkyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₃ alkyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₂ alkyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₁ alkyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₀ alkyl.

In some embodiments, R⁷ is optionally substituted C₁₀₋₅₀ alkyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₄₀ alkyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₃₀ alkyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₂₀ alkyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₉ alkyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₈ alkyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₇ alkyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₆ alkyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₅ alkyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₄ alkyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₃ alkyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₂ alkyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₁ alkyl.

In some embodiments, R⁷ is optionally substituted C₁₁₋₅₀ alkyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₄₀ alkyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₃₀ alkyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₂₀ alkyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₉ alkyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₈alkyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₇ alkyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₆ alkyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₅ alkyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₄ alkyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₃ alkyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₂ alkyl.

In some embodiments, R⁷ is optionally substituted C₁₂₋₅₀ alkyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₄₀ alkyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₃₀ alkyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₂₀ alkyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₉ alkyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₈ alkyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₇ alkyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₆ alkyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₅ alkyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₄ alkyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₃ alkyl.

In some embodiments, R⁷ is optionally substituted C₆ alkyl. In some embodiments, R⁷ is optionally substituted C₇ alkyl. In some embodiments, R⁷ is optionally substituted C₈ alkyl. In some embodiments, R⁷ is optionally substituted C₉ alkyl. In some embodiments, R⁷ is optionally substituted C₁₀alkyl. In some embodiments, R⁷ is optionally substituted C₁₁ alkyl. In some embodiments, R⁷ is optionally substituted C₁₂ alkyl. In some embodiments, R⁷ is optionally substituted C₁₃ alkyl. In some embodiments, R⁷ is optionally substituted C₁₄ alkyl. In some embodiments, R⁷ is optionally substituted C₁₅ alkyl. In some embodiments, R⁷ is optionally substituted C₁₆ alkyl. In some embodiments, R⁷ is optionally substituted C₁₇ alkyl. In some embodiments, R⁷ is optionally substituted C₁₈ alkyl. In some embodiments, R⁷ is optionally substituted C₁₉ alkyl. In some embodiments, R⁷ is optionally substituted C₂₀ alkyl.

In some embodiments, for example, in any of the above embodiments, R⁷ is a substituted alkyl group. In some embodiments, R⁷ is an unsubstituted alkyl group. In some embodiments, R⁷ is an optionally substituted straight-chain alkyl group. In some embodiments, R⁷ is a substituted straight-chain alkyl group. In some embodiments, R⁷ is an unsubstituted straight-chain alkyl group. In some embodiments, R⁷ is an optionally substituted branched alkyl group. In some embodiments, R⁷ is a substituted branched alkyl group. In some embodiments, R⁷ is an unsubstituted branched alkyl group.

In some embodiments, R⁷ is optionally substituted alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₅₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₄₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₃₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₂₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₉ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₈ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₇ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₆ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₅ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₄ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₃ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₂ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₁ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₉ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₈ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₇ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₋₆ alkenyl.

In some embodiments, R⁷ is optionally substituted C₄₋₅₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₄₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₃₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₂₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₉ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₈ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₇ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₆ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₅ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₄ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₃ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₂ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₁ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₉ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₈ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₇ alkenyl. In some embodiments, R⁷ is optionally substituted C₄₋₆ alkenyl.

In some embodiments, R⁷ is optionally substituted C₆₋₅₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₄₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₃₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₂₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₉ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₈ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₇ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₆ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₅ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₄ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₃ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₂ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₁ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₉ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₈ alkenyl. In some embodiments, R⁷ is optionally substituted C₆₋₇ alkenyl.

In some embodiments, R⁷ is optionally substituted C₈₋₅₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₈₋₄₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₈₋₃₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₈₋₂₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₉ alkenyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₈ alkenyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₇ alkenyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₆ alkenyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₅ alkenyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₄ alkenyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₃ alkenyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₂ alkenyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₁ alkenyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₈₋₉ alkenyl.

In some embodiments, R⁷ is optionally substituted C₉₋₅₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₉₋₄₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₉₋₃₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₉₋₂₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₉ alkenyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₈ alkenyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₇ alkenyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₆ alkenyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₅ alkenyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₄ alkenyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₃ alkenyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₂ alkenyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₁ alkenyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₀ alkenyl.

In some embodiments, R⁷ is optionally substituted C₁₀₋₅₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₄₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₃₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₂₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₉ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₈ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₇ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₆ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₅ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₄ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₃ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₂ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₁ alkenyl.

In some embodiments, R⁷ is optionally substituted C₁₁₋₅₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₄₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₃₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₂₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₉ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₁ s alkenyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₇ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₆ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₅ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₄ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₃ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₂ alkenyl.

In some embodiments, R⁷ is optionally substituted C₁₂₋₅₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₄₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₃₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₂₀ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₉ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₈ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₇ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₆ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₅ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₄ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₃ alkenyl.

In some embodiments, R⁷ is optionally substituted C₆ alkenyl. In some embodiments, R⁷ is optionally substituted C₇ alkenyl. In some embodiments, R⁷ is optionally substituted C₈ alkenyl. In some embodiments, R⁷ is optionally substituted C₉ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₀alkenyl. In some embodiments, R⁷ is optionally substituted C₁₁ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₂ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₃ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₄ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₅ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₆ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₇ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₈ alkenyl. In some embodiments, R⁷ is optionally substituted C₁₉ alkenyl. In some embodiments, R⁷ is optionally substituted C₂₀ alkenyl.

In some embodiments, for example, in any of the above embodiments, R⁷ is a substituted alkenyl group. In some embodiments, R⁷ is an unsubstituted alkenyl group. In some embodiments, R⁷ is an optionally substituted straight-chain alkenyl group. In some embodiments, R⁷ is a substituted straight-chain alkenyl group. In some embodiments, R⁷ is an unsubstituted straight-chain alkenyl group. In some embodiments, R⁷ is an optionally substituted branched alkenyl group. In some embodiments, R⁷ is a substituted branched alkenyl group. In some embodiments, R⁷ is an unsubstituted branched alkenyl group.

In some embodiments, R⁷ is optionally substituted alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₅₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₄₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₃₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₂₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₉ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₈ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₇ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₆ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₅ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₄ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₃ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₂ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₁ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₁₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₉ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₈ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₇ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₋₆ alkynyl.

In some embodiments, R⁷ is optionally substituted C₄₋₅₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₄₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₃₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₂₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₉ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₈ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₇ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₆ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₅ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₄ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₃ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₂ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₁ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₁₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₉ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₈ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₇ alkynyl. In some embodiments, R⁷ is optionally substituted C₄₋₆ alkynyl.

In some embodiments, R⁷ is optionally substituted C₆₋₅₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₄₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₃₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₂₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₉ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₈ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₇ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₆ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₅ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₄ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₃ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₂ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₁ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₁₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₉ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₈ alkynyl. In some embodiments, R⁷ is optionally substituted C₆₋₇ alkynyl.

In some embodiments, R⁷ is optionally substituted C₈₋₅₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₈₋₄₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₈₋₃₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₈₋₂₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₉ alkynyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₈ alkynyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₇ alkynyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₆ alkynyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₅ alkynyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₄ alkynyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₃ alkynyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₂ alkynyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₁ alkynyl. In some embodiments, R⁷ is optionally substituted C₈₋₁₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₈₋₉ alkynyl.

In some embodiments, R⁷ is optionally substituted C₉₋₅₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₉₋₄₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₉₋₃₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₉₋₂₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₉ alkynyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₈ alkynyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₇ alkynyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₆ alkynyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₅ alkynyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₄ alkynyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₃ alkynyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₂ alkynyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₁ alkynyl. In some embodiments, R⁷ is optionally substituted C₉₋₁₀ alkynyl.

In some embodiments, R⁷ is optionally substituted C₁₀₋₅₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₄₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₃₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₂₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₉ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₈ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₇ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₆ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₅ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₄ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₃ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₂ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₀₋₁₁ alkynyl.

In some embodiments, R⁷ is optionally substituted C₁₁₋₅₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₄₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₃₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₂₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₉ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₁ s alkynyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₇ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₆ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₅ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₄ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₃ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₁₋₁₂ alkynyl.

In some embodiments, R⁷ is optionally substituted C₁₂₋₅₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₄₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₃₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₂₀ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₉ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₈ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₇ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₆ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₅ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₄ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₂₋₁₃ alkynyl.

In some embodiments, R⁷ is optionally substituted C₆ alkynyl. In some embodiments, R⁷ is optionally substituted C₇ alkynyl. In some embodiments, R⁷ is optionally substituted C₈ alkynyl. In some embodiments, R⁷ is optionally substituted C₉ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₀alkynyl. In some embodiments, R⁷ is optionally substituted C₁₁ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₂ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₃ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₄ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₅ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₆ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₇ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₈ alkynyl. In some embodiments, R⁷ is optionally substituted C₁₉ alkynyl. In some embodiments, R⁷ is optionally substituted C₂₀ alkynyl.

In some embodiments, for example, in any of the above embodiments, R⁷ is a substituted alkynyl group. In some embodiments, R is an unsubstituted alkynyl group. In some embodiments, R⁷ is an optionally substituted straight-chain alkynyl group. In some embodiments, R is a substituted straight-chain alkynyl group. In some embodiments, R⁷ is an unsubstituted straight-chain alkynyl group. In some embodiments, R is an optionally substituted branched alkynyl group. In some embodiments, R⁷ is a substituted branched alkynyl group. In some embodiments, R⁷ is an unsubstituted branched alkynyl group.

In some embodiments, R⁷ is optionally substituted carbocyclyl. In some embodiments, R⁷ is optionally substituted heterocyclyl. In some embodiments, R⁷ is optionally substituted aryl. In some embodiments, R⁷ is optionally substituted heteroaryl. In some embodiments, R⁷ is a nitrogen protecting group.

In some embodiments, R⁷ is a group of formula (i). In some embodiments, R⁷ is a group of formula (i-a). In some embodiments, R⁷ is a group of formula

In some embodiments, R⁷ is a group of formula (i-b). In some embodiments, R⁷ is a group of formula (ii). In some embodiments, R⁷ is a group of formula (iii).

In some embodiments, at least one instance of R⁶ and R⁷ is a group of the formula (i), (ii) or (iii). In some embodiments, each instance of R⁶ and R⁷ is independently a group of the formula (i), (ii) or (iii). In some embodiments, each instance of R⁶ and R⁷ is independently a group of the formula (i). In some embodiments, each instance of R⁶ and R⁷ is independently a group of the formula (i-a). In some embodiments, each instance of R⁶ and R⁷ is independently a group of the formula (i-b). In some embodiments, each instance of R⁶ and R⁷ is independently a group of the formula (ii). In some embodiments, each instance of R⁶ and R⁷ is independently a group of the formula (iii).

In some embodiments, R⁶ and R⁷ are the same. In some embodiments, R⁶ and R⁷ are different.

In certain embodiments, both R⁶ and R⁷ are hydrogen. In certain embodiments, R⁶ is hydrogen and R⁷ is a group of the formula (i), (ii), or (iii). In certain embodiments, R⁶ is hydrogen and R⁷ is a group of the formula (i). In certain embodiments, R⁶ is hydrogen and R⁷ is a group of the formula (ii). In certain embodiments, R⁶ is hydrogen and R⁷ is a group of the formula (iii). In certain embodiments, each of R⁶ and R⁷ is independently a group of the formula (i), (ii), or (iii). In certain embodiments, each of R⁶ and R⁷ is independently a group of the formula (i). In certain embodiments, each of R⁶ and R⁷ is independently a group of the formula (ii). In certain embodiments, each of R⁶ and R⁷ is independently a group of the formula (iii). In certain embodiments, R⁶ and R⁷ are the same group, which is selected from formulas (i), (ii), and (iii). In some embodiments, R⁶ and R⁷ are the same group of formula (i). In some embodiments, R⁶ and R⁷ are the same group of formula (i-a). In some embodiments, R⁶ and R⁷ are the same group of formula (i-a1). In some embodiments, R⁶ and R⁷ are the same group of formula (i-b).

In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is as defined above and described herein. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₁₋₅₀alkyl, optionally substituted C₂₋₅₀alkenyl, optionally substituted C₂₋₅₀alkynyl, optionally substituted heteroC₁₋₅₀alkyl, optionally substituted heteroC₂₋₅₀alkenyl, or optionally substituted heteroC₂₋₅₀alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₈₋₅₀alkyl, optionally substituted C₈₋₅₀alkenyl, optionally substituted C₈₋₅₀alkynyl, optionally substituted heteroC₁₋₅₀alkyl, optionally substituted heteroC₅₋₅₀alkenyl, or optionally substituted heteroC₅₋₅₀alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₅₋₄₀alkyl, optionally substituted C₅₋₄₀alkenyl, optionally substituted C₅₋₄₀alkynyl, optionally substituted heteroC₅₋₄₀alkyl, optionally substituted heteroC₅₋₄₀alkenyl, or optionally substituted heteroC₅₋₄₀alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₅₋₃₀alkyl, optionally substituted C₅₋₃₀alkenyl, optionally substituted C₅₋₃₀alkynyl, optionally substituted heteroC₅₋₃₀alkyl, optionally substituted heteroC₅₋₃₀alkenyl, or optionally substituted heteroC₅₋₃₀alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₅₋₂₅alkyl, optionally substituted C₅₋₂₅alkenyl, optionally substituted C₅₋₂₅alkynyl, optionally substituted heteroC₅₋₂₅alkyl, optionally substituted heteroC₅₋₂₅alkenyl, or optionally substituted heteroC₈₋₂₅alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₅₋₂₀alkyl, optionally substituted C₅₋₂₀alkenyl, optionally substituted C₅₋₂₀alkynyl, optionally substituted heteroC₅₋₂₀alkyl, optionally substituted heteroC₅₋₂₀alkenyl, or optionally substituted heteroC₅₋₂₀alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₅₋₁₅alkyl, optionally substituted C₅₋₁₅alkenyl, optionally substituted C₅₋₁₅alkynyl, optionally substituted heteroC₅₋₁₅alkyl, optionally substituted heteroC₈₋₁₅alkenyl, or optionally substituted heteroC₈₋₁₅alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₅ alkyl, optionally substituted C₅ alkenyl, optionally substituted C₅ alkynyl, optionally substituted heteroC₅alkyl, optionally substituted heteroC₅alkenyl, or optionally substituted heteroC₅alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₆ alkyl, optionally substituted C₆ alkenyl, optionally substituted C₆ alkynyl, optionally substituted heteroC₆alkyl, optionally substituted heteroC₆alkenyl, or optionally substituted heteroC₆alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₇ alkyl, optionally substituted C₇ alkenyl, optionally substituted C₇ alkynyl, optionally substituted heteroC₇alkyl, optionally substituted heteroC₇alkenyl, or optionally substituted heteroC₇alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₈ alkyl, optionally substituted C₈ alkenyl, optionally substituted C₈ alkynyl, optionally substituted heteroC₈alkyl, optionally substituted heteroC₈alkenyl, or optionally substituted heteroC₈alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₉ alkyl, optionally substituted C₉ alkenyl, optionally substituted C₉ alkynyl, optionally substituted heteroC₉alkyl, optionally substituted heteroC₉alkenyl, or optionally substituted heteroC₉alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R is optionally substituted C₁₀alkyl, optionally substituted C₁₀alkenyl, optionally substituted C₁₀ alkynyl, optionally substituted heteroC₁₀alkyl, optionally substituted heteroC₁₀alkenyl, or optionally substituted heteroC₁₀alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₁₁ alkyl, optionally substituted C₁₁ alkenyl, optionally substituted C₁₁ alkynyl, optionally substituted heteroC₁₁alkyl, optionally substituted heteroC₁₁alkenyl, or optionally substituted heteroC₁₁alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₁₂ alkyl, optionally substituted C₁₂ alkenyl, optionally substituted C₁₂ alkynyl, optionally substituted heteroC₁₂alkyl, optionally substituted heteroC₁₂alkenyl, or optionally substituted heteroC₁₂alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₁₃ alkyl, optionally substituted C₁₃ alkenyl, optionally substituted C₁₃ alkynyl, optionally substituted heteroC₁₃alkyl, optionally substituted heteroC₁₃alkenyl, or optionally substituted heteroC₁₃alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₁₄ alkyl, optionally substituted C₁₄ alkenyl, optionally substituted C₁₄ alkynyl, optionally substituted heteroC₁₄alkyl, optionally substituted heteroC₁₄alkenyl, or optionally substituted heteroC₁₄alkynyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₁₅ alkyl, optionally substituted C₁₅ alkenyl, optionally substituted C₁₅ alkynyl, optionally substituted heteroC₁₅alkyl, optionally substituted heteroC₁₅alkenyl, or optionally substituted heteroC₁₅alkynyl.

In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is as defined above and described herein. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₁₋₅₀alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₅₋₅₀alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₅₋₄₀alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₅₋₃₀alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₈₋₂₅alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₅₋₂₀alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₅₋₁₅alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₅ alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₆ alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₇ alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₈ alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₉ alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₁₀alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₁₁ alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₁₂ alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₁₃ alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₁₄ alkyl. In some embodiments, R⁶ and R⁷ are the same group of formula

wherein R^(L) is optionally substituted C₁₅ alkyl.

As generally defined above, each occurrence of R^(A1) is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, an oxygen protecting group when attached to an oxygen atom, a sulfur protecting group when attached to an sulfur atom, a nitrogen protecting group when attached to a nitrogen atom, or two R^(A1) groups, together with the nitrogen atom to which they are attached, are joined to form an optionally substituted heterocyclic or optionally substituted heteroaryl ring.

In some embodiments, R^(A1) is hydrogen. In some embodiments, R^(A1) is optionally substituted alkyl. In some embodiments, R^(A1) is optionally substituted alkenyl. In some embodiments, R^(A1) is optionally substituted alkynyl. In some embodiments, R^(A1) is optionally substituted carbocyclyl. In some embodiments, R^(A1) is optionally substituted heterocyclyl. In some embodiments, R^(A1) is optionally substituted aryl. In some embodiments, R^(A1) is optionally substituted heteroaryl. In some embodiments, R^(A1) is an oxygen protecting group when attached to an oxygen atom. In some embodiments, R^(A1) is a sulfur protecting group when attached to a sulfur atom. In some embodiments, R^(A1) is a nitrogen protecting group when attached to a nitrogen atom. In some embodiments, two R^(A1) groups, together with the nitrogen atom to which they are attached, are joined to form an optionally substituted heterocyclic or optionally substituted heteroaryl ring.

As generally defined above, each instance of R′ is independently hydrogen or optionally substituted alkyl. In some embodiments, R′ is hydrogen. In some embodiments, R′ is substituted alkyl. In certain embodiments, at least one instance of R′ is hydrogen. In certain embodiments, at least two instances of R′ is hydrogen. In certain embodiments, each instance of R′ is hydrogen. In certain embodiments, at least one instance of R′ is optionally substituted alkyl, e.g., methyl. In certain embodiments, at least two instances of R′ is optionally substituted alkyl, e.g., methyl. In some embodiments, at least one instance of R′ is hydrogen, and at least one instance of R′ is optionally substituted alkyl. In certain embodiments, one instance of R′ is optionally substituted alkyl, and the rest are hydrogen.

As generally defined above, X is O, S, or NR^(X). In some embodiments, X is O. In some embodiments, X is S. In some embodiments, X is NR^(X), wherein R^(X) is as defined above and described herein.

As generally defined above, R^(X) is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or a nitrogen protecting group. In some embodiments, R^(X) is hydrogen. In some embodiments, R^(X) is optionally substituted alkyl. In some embodiments, R^(X) is optionally substituted alkenyl. In some embodiments, R^(X) is optionally substituted alkynyl. In some embodiments, R^(X) is optionally substituted carbocyclyl. In some embodiments, R^(X) is optionally substituted heterocyclyl. In some embodiments, R^(X) is optionally substituted aryl. In some embodiments, R^(X) is optionally substituted heteroaryl. In some embodiments, R^(X) is a nitrogen protecting group.

As generally defined above, Y is O, S, or NR^(Y). In some embodiments, Y is O. In some embodiments, Y is S. In some embodiments, Y is NR^(Y), wherein R^(Y) is as defined above and described herein.

As generally defined above, R^(Y) is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or a nitrogen protecting group. In some embodiments, R^(Y) is hydrogen. In some embodiments, R^(Y) is optionally substituted alkyl. In some embodiments, R^(Y) is optionally substituted alkenyl. In some embodiments, R^(Y) is optionally substituted alkynyl. In some embodiments, R^(Y) is optionally substituted carbocyclyl. In some embodiments, R^(Y) is optionally substituted heterocyclyl. In some embodiments, R^(Y) is optionally substituted aryl. In some embodiments, R^(Y) is optionally substituted heteroaryl. In some embodiments, R^(Y) is a nitrogen protecting group.

As generally defined above, R^(P) is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, an oxygen protecting group when attached to an oxygen atom, a sulfur protecting group when attached to a sulfur atom, or a nitrogen protecting group when attached to a nitrogen atom. In some embodiments, R^(P) is hydrogen. In some embodiments, R^(P) is optionally substituted alkyl. In some embodiments, R^(P) is optionally substituted alkenyl. In some embodiments, R^(P) is optionally substituted alkynyl. In some embodiments, R^(P) is optionally substituted carbocyclyl. In some embodiments, R^(P) is optionally substituted heterocyclyl. In some embodiments, R^(P) is optionally substituted aryl. In some embodiments, R^(P) is optionally substituted heteroaryl. In some embodiments, R^(P) is an oxygen protecting group when attached to an oxygen atom. In some embodiments, R^(P) is a sulfur protecting group when attached to a sulfur atom. In some embodiments, R^(P) is a nitrogen protecting group when attached to a nitrogen atom.

As generally defined above, R^(L) is optionally substituted C₁₋₅₀ alkyl, optionally substituted C₂₋₅₀ alkenyl, optionally substituted C₂₋₅₀ alkynyl, optionally substituted heteroC₁₋₅₀ alkyl, optionally substituted heteroC₂₋₅₀ alkenyl, optionally substituted heteroC₂₋₅₀ alkynyl, or a polymer.

In some embodiments, R^(L) is optionally substituted C₁₋₅₀ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₅₀ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₄₀ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₃₀ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₂₀ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₉ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₈ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₇ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₆ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₅ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₄ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₃ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₂ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₁ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₀ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₉ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₈ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₇ alkyl. In some embodiments, R^(L) is optionally substituted C₂₋₆ alkyl.

In some embodiments, R^(L) is optionally substituted C₄₋₅₀ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₄₀ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₃₀ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₂₀ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₉ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₈ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₇ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₆ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₅ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₄ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₃ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₂ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₁ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₀ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₉ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₈ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₇ alkyl. In some embodiments, R^(L) is optionally substituted C₄₋₆ alkyl.

In some embodiments, R^(L) is optionally substituted C₆₋₅₀ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₄₀ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₃₀ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₂₀ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₉ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₈ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₇ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₆ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₅ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₄ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₃ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₂ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₁ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₀ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₉ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₈ alkyl. In some embodiments, R^(L) is optionally substituted C₆₋₇ alkyl.

In some embodiments, R^(L) is optionally substituted C₈₋₅₀ alkyl. In some embodiments, R^(L) is optionally substituted C₈₋₄₀ alkyl. In some embodiments, R^(L) is optionally substituted C₈₋₃₀ alkyl. In some embodiments, R^(L) is optionally substituted C₈₋₂₀ alkyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₉ alkyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₅ alkyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₇ alkyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₆ alkyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₅ alkyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₄ alkyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₃ alkyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₂ alkyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₁ alkyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₀ alkyl. In some embodiments, R^(L) is optionally substituted C₈₋₉ alkyl.

In some embodiments, R^(L) is optionally substituted C₉₋₅₀ alkyl. In some embodiments, R^(L) is optionally substituted C₉₋₄₀ alkyl. In some embodiments, R^(L) is optionally substituted C₉₋₃₀ alkyl. In some embodiments, R^(L) is optionally substituted C₉₋₂₀ alkyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₉ alkyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₈ alkyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₇ alkyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₆ alkyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₅ alkyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₄ alkyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₃ alkyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₂ alkyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₁ alkyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₀ alkyl.

In some embodiments, R^(L) is optionally substituted C₁₀₋₅₀ alkyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₄₀ alkyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₃₀ alkyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₂₀ alkyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₉ alkyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₅ alkyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₇ alkyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₆ alkyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₅ alkyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₄ alkyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₃ alkyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₂ alkyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₁ alkyl.

In some embodiments, R^(L) is optionally substituted C₁₁₋₅₀ alkyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₄₀ alkyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₃₀ alkyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₂₀ alkyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₉ alkyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₈alkyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₇ alkyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₆ alkyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₅ alkyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₄ alkyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₃ alkyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₂ alkyl.

In some embodiments, R^(L) is optionally substituted C₁₂₋₅₀ alkyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₄₀ alkyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₃₀ alkyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₂₀ alkyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₉ alkyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₈ alkyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₇ alkyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₆ alkyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₅ alkyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₄ alkyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₃ alkyl.

In some embodiments, R^(L) is optionally substituted C₆ alkyl. In some embodiments, R^(L) is optionally substituted C₇ alkyl. In some embodiments, R^(L) is optionally substituted C₈ alkyl. In some embodiments, R^(L) is optionally substituted C₉ alkyl. In some embodiments, R^(L) is optionally substituted C₁₀alkyl. In some embodiments, R^(L) is optionally substituted C₁₁ alkyl. In some embodiments, R^(L) is optionally substituted C₁₂ alkyl. In some embodiments, R^(L) is optionally substituted C₁₃ alkyl. In some embodiments, R^(L) is optionally substituted C₁₄ alkyl. In some embodiments, R^(L) is optionally substituted C₁₅ alkyl. In some embodiments, R^(L) is optionally substituted C₁₆ alkyl. In some embodiments, R^(L) is optionally substituted C₁₇ alkyl. In some embodiments, R^(L) is optionally substituted C₁₈ alkyl. In some embodiments, R^(L) is optionally substituted C₁₉ alkyl. In some embodiments, R^(L) is optionally substituted C₂₀ alkyl.

In some embodiments, for example, in any of the above embodiments, R^(L) is a substituted alkyl group. In some embodiments, R^(L) is an unsubstituted alkyl group. In some embodiments, R^(L) is an optionally substituted straight-chain alkyl group. In some embodiments, R^(L) is a substituted straight-chain alkyl group. In some embodiments, R^(L) is an unsubstituted straight-chain alkyl group. In some embodiments, R^(L) is an optionally substituted branched alkyl group. In some embodiments, R^(L) is a substituted branched alkyl group. In some embodiments, R^(L) is an unsubstituted branched alkyl group.

In certain embodiments, at least one instance of R^(L) is an unsubstituted alkyl. Exemplary unsubstituted alkyl groups include, but are not limited to, —CH₃, —C₂H₅, —C₃H₇, —C₄H₉, —C₅H₁₁, —C₆H₁₃, —C₇H₁₅, —C₈H₁₇, —C₉H₁₉, —C₁₀H₂₁, —C₁₁H₂₃, —C₁₂H₂₅, —C₁₃H₂₇, —C₁₄H₂₉, —C₁₅H₃₁, —C₁₆H₃₃, —C₁₇H₃₅, —C₁₈H₃₇, —C₁₉H₃₉, —C₂₀H₄₁, —C₂₁H₄₃, —C₂₂H₄₅, —C₂₃H₄₇, —C₂₄H₄₉, and —C₂₅H₅₁.

In certain embodiments, at least one instance of R^(L) is a substituted alkyl. For example, in certain embodiments, at least one instance of R^(L) is an alkyl substituted with one or more fluorine substituents. Exemplary fluorinated alkyl groups include, but are not limited to:

In some embodiments, R^(L) is optionally substituted C₂₋₅₀ alkenyl. In some some embodiments, R^(L) is optionally substituted C₂₋₁₉ alkenyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₈ alkenyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₇ alkenyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₆ alkenyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₅ alkenyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₄ alkenyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₃ alkenyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₂ alkenyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₁ alkenyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₂₋₉ alkenyl. In some embodiments, R^(L) is optionally substituted C₂₋₈ alkenyl. In some embodiments, R^(L) is optionally substituted C₂₋₇ alkenyl. In some embodiments, R^(L) is optionally substituted C₂₋₆ alkenyl.

In some embodiments, R^(L) is optionally substituted C₄₋₅₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₄₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₃₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₂₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₉ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₈ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₇ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₆ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₅ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₄ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₃ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₂ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₁ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₉ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₈ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₇ alkenyl. In some embodiments, R^(L) is optionally substituted C₄₋₆ alkenyl.

In some embodiments, R^(L) is optionally substituted C₆₋₅₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₄₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₃₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₂₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₉ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₈ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₇ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₆ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₅ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₄ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₃ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₂ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₁ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₉ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₈ alkenyl. In some embodiments, R^(L) is optionally substituted C₆₋₇ alkenyl.

In some embodiments, R^(L) is optionally substituted C₈₋₅₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₈₋₄₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₈₋₃₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₈₋₂₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₉ alkenyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₈ alkenyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₇ alkenyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₆ alkenyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₅ alkenyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₄ alkenyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₃ alkenyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₂ alkenyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₁ alkenyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₈₋₉ alkenyl.

In some embodiments, R^(L) is optionally substituted C₉₋₅₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₉₋₄₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₉₋₃₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₉₋₂₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₉ alkenyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₈ alkenyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₇ alkenyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₆ alkenyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₅ alkenyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₄ alkenyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₃ alkenyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₂ alkenyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₁ alkenyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₀ alkenyl.

In some embodiments, R^(L) is optionally substituted C₁₀₋₅₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₄₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₃₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₂₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₉ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₈ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₇ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₆ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₅ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₄ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₃ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₂ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₁ alkenyl.

In some embodiments, R^(L) is optionally substituted C₁₁₋₅₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₄₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₃₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₂₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₉ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₈ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₇ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₆ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₅ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₄ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₃ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₂ alkenyl.

In some embodiments, R^(L) is optionally substituted C₁₂₋₅₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₄₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₃₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₂₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₉ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₈ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₇ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₆ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₅ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₄ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₃ alkenyl.

In some embodiments, R^(L) is optionally substituted C₆ alkenyl. In some embodiments, R^(L) is optionally substituted C₇ alkenyl. In some embodiments, R^(L) is optionally substituted C₈ alkenyl. In some embodiments, R^(L) is optionally substituted C₉ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₀ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₁ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₂ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₃ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₄ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₅ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₆ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₇ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₈ alkenyl. In some embodiments, R^(L) is optionally substituted C₁₉ alkenyl. In some embodiments, R^(L) is optionally substituted C₂₀ alkenyl.

In some embodiments, for example, in any of the above embodiments, R^(L) is a substituted alkyl group. In some embodiments, R^(L) is an unsubstituted alkyl group. In some embodiments, R^(L) is an optionally substituted straight-chain alkenyl group. In some embodiments, R^(L) is a substituted straight-chain alkenyl group. In some embodiments, R^(L) is an unsubstituted straight-chain alkenyl group. In some embodiments, R^(L) is an optionally substituted branched alkenyl group. In some embodiments, R^(L) is a substituted branched alkenyl group. In some embodiments, R^(L) is an unsubstituted branched alkenyl group.

Exemplary unsubstituted alkenyl group include, but are not limited to:

-   Mylistoleic —(CH₂)₂CH═CH(C_(H2))CH₃, -   Palmitoliec —(CH₂)₇CH═CH(CH₂)₅CH₃, -   Sapienic —(CH₂)₄CH═CH(CH₂)₈CH₃; -   Oleic —(CH₂)₇CH═CH(CH₂)₇CH₃, -   Linoleic —(CH₂)₇CH═CHCH₂CH═CH(CH₂)₄CH₃, -   α-Linolenic —(CH₂)₇CH═CHCH₂CH═CHCH₂CH═CHCH₂CH₃, -   Arachinodonic —(CH)₂)₃CH═CHCH₂CH═CHCH₂CHCH₂CH═CHCH₂CH═CH(CH₂)₄CH₃, -   Eicosapentaenoic     —(CH₂)₃CH═CHCH₂CH═CHCH₂CH═CHCH₂CH═CHCH₂CH═CHCH₂CH═CHCH₂CH₃, -   Erucic —(CH₂)₁₁CH═CH(CH₂)₇CH₃, and -   Docosahexaenoic     —(CH₂)₂CH═CHCH₂CH═CHCH₂CH═CHCH₂CH═CHCH₂CH═CHCH₂CH═CH—CH₂CH₃.

In some embodiments, wherein R^(L) is defined as a C₆₋₅₀alkyl or C₆₋₅₀alkenyl groups, such groups are meant to encompass lipophilic groups (also referred to as a “lipid tail”). Lipophilic groups comprise a group of molecules that include fats, waxes, oils, fatty acids, and the like. Lipid tails present in these lipid groups can be saturated and unsaturated, depending on whether or not the lipid tail comprises double bonds. The lipid tail can also comprise different lengths, often categorized as medium (i.e., with tails between 7-12 carbons, e.g., C₇₋₁₂ alkyl or C₇₋₁₂ alkenyl), long (i.e., with tails greater than 12 carbons and up to 22 carbons, e.g., C₁₃₋₂₂alkyl or C₁₃₋₂₂ alkenyl), or very long (i.e., with tails greater than 22 carbons, e.g., C₂₃₋₃₀ alkyl or C₂₃₋₃₀ alkenyl).

In some embodiments, R^(L) is optionally substituted C₂₋₅₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₄₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₃₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₂₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₉ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₈ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₇ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₆ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₅ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₄ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₃ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₂ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₁ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₁₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₉ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₈ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₇ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₋₆ alkynyl.

In some embodiments, R^(L) is optionally substituted C₄₋₅₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₄₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₃₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₂₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₉ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₈ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₇ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₆ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₅ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₄ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₃ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₂ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₁ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₁₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₉ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₈ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₇ alkynyl. In some embodiments, R^(L) is optionally substituted C₄₋₆ alkynyl.

In some embodiments, R^(L) is optionally substituted C₆₋₅₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₄₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₃₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₂₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₉ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₈ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₇ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₆ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₅ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₄ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₃ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₂ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₁ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₁₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₉ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₈ alkynyl. In some embodiments, R^(L) is optionally substituted C₆₋₇ alkynyl.

In some embodiments, R^(L) is optionally substituted C₈₋₅₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₈₋₄₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₈₋₃₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₈₋₂₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₉ alkynyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₈ alkynyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₇ alkynyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₆ alkynyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₅ alkynyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₄ alkynyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₃ alkynyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₂ alkynyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₁ alkynyl. In some embodiments, R^(L) is optionally substituted C₈₋₁₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₈₋₉ alkynyl.

In some embodiments, R^(L) is optionally substituted C₉₋₅₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₉₋₄₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₉₋₃₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₉₋₂₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₉ alkynyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₈ alkynyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₇ alkynyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₆ alkynyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₅ alkynyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₄ alkynyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₃ alkynyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₂ alkynyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₁ alkynyl. In some embodiments, R^(L) is optionally substituted C₉₋₁₀ alkynyl.

In some embodiments, R^(L) is optionally substituted C₁₀₋₅₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₄₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₃₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₂₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₉ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₈ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₇ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₆ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₅ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₄ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₃ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₂ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₀₋₁₁ alkynyl.

In some embodiments, R^(L) is optionally substituted C₁₁₋₅₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₄₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₃₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₂₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₉ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₈ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₇ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₆ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₅ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₄ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₃ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₁₋₁₂ alkynyl.

In some embodiments, R^(L) is optionally substituted C₁₂₋₅₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₄₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₃₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₂₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₉ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₈ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₇ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₆ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₅ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₄ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₂₋₁₃ alkynyl.

In some embodiments, R^(L) is optionally substituted C₆ alkynyl. In some embodiments, R^(L) is optionally substituted C₇ alkynyl. In some embodiments, R^(L) is optionally substituted C₈ alkynyl. In some embodiments, R^(L) is optionally substituted C₉ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₀ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₁ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₂ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₃ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₄ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₅ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₆ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₇ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₈ alkynyl. In some embodiments, R^(L) is optionally substituted C₁₉ alkynyl. In some embodiments, R^(L) is optionally substituted C₂₀ alkynyl.

In some embodiments, for example, in any of the above embodiments, R^(L) is a substituted alkynyl group. In some embodiments, R^(L) is an unsubstituted alkynyl group. In some embodiments, R^(L) is an optionally substituted straight-chain alkyl group. In some embodiments, R^(L) is an optionally substituted straight-chain alkynyl group. In some embodiments, R^(L) is a substituted straight-chain alkynyl group. In some embodiments, R^(L) is an unsubstituted straight-chain alkynyl group. In some embodiments, R^(L) is an optionally substituted branched alkynyl group. In some embodiments, R^(L) is a substituted branched alkynyl group. In some embodiments, R^(L) is an unsubstituted branched alkynyl group.

In some embodiments, R^(L) is optionally substituted heteroC₁₋₅₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₅₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₄₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₃₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₂₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₉alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₈alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₇alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₆alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₅alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₄alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₃alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₂alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₁alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₉alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₅alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₇alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₆alkyl.

In some embodiments, R^(L) is optionally substituted heteroC₄₋₅₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₄₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₃₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₂₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₉alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₈alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₇alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₆alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₅alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₄alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₃alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₂alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₁alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₉alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₈alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₇alkyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₆alkyl.

In some embodiments, R^(L) is optionally substituted heteroC₆₋₅₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₄₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₃₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₂₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₉alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₈alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₇alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₆alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₅alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₄alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₃alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₂alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₁alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₉alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₈alkyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₇alkyl.

In some embodiments, R^(L) is optionally substituted heteroC₈₋₅₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₋₄₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₃₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₂₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₋₁₉alkyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₅alkyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₇alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₋₁₆alkyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₅alkyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₄alkyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₃alkyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₂alkyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₁alkyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₉alkyl.

In some embodiments, R^(L) is optionally substituted heteroC₉₋₅₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₄₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₃₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₂₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₉alkyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₈alkyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₇alkyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₆alkyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₅alkyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₄alkyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₃alkyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₂alkyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₁alkyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₀alkyl.

In some embodiments, R^(L) is optionally substituted heteroC₁₋₅₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₄₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₃₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₋₂₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₉alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₈alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₇alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₆alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₅alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₄alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₃alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₂alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₁alkyl.

In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₅₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₄₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₃₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₂₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₉alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₈alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₇alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₆alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₅alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₄alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₃alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₂alkyl.

In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₅₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₄₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₃₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₂₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₉alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₈alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₇alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₆alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₅alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₄alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₃alkyl.

In some embodiments, R^(L) is optionally substituted heteroC₆alkyl. In some embodiments, R^(L) is optionally substituted heteroC₇alkyl. In some embodiments, R^(L) is optionally substituted heteroC₅alkyl. In some embodiments, R^(L) is optionally substituted heteroC₉alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₃alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₄alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₅alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₆alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₇alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₅alkyl. In some embodiments, R^(L) is optionally substituted heteroC₁₉alkyl. In some embodiments, R^(L) is optionally substituted heteroC₂₀alkyl.

In some embodiments, for example, in any of the above embodiments, R^(L) is a substituted heteroalkyl group. In some embodiments, R^(L) is an unsubstituted heteroalkyl group. In some embodiments, R^(L) is an optionally substituted straight-chain heteroalkyl group. In some embodiments, R^(L) is a substituted straight-chain heteroalkyl group. In some embodiments, R^(L) is an unsubstituted straight-chain heteroalkyl group. In some embodiments, R^(L) is an optionally substituted branched heteroalkyl group. In some embodiments, R^(L) is a substituted branched heteroalkyl group. In some embodiments, R^(L) is an unsubstituted branched heteroalkyl group.

Exemplary unsubstituted heteroalkyl groups include, but are not limited to:

In some embodiments, R^(L) is optionally substituted heteroC₂₋₅₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₄₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₃₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₂₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₉alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₈alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₇alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₆alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₄alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₃alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₂alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₁alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₉alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₇alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₆alkenyl.

In some embodiments, R^(L) is optionally substituted heteroC₄₋₅₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₄₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₃₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₂₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₉alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₇alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₆alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₄alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₃alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₂alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₁alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₉alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₇alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₆alkenyl.

In some embodiments, R^(L) is optionally substituted heteroC₆₋₅₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₄₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₃₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₂₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₉alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₈alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₇alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₆alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₄alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₃alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₂alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₁alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₉alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₈alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₇alkenyl.

In some embodiments, R^(L) is optionally substituted heteroC₈₋₅₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₄₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₅₋₃₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₂₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₉alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₅₋₁₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₇alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₆alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₄alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₃alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₂alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₁alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₉alkenyl.

In some embodiments, R^(L) is optionally substituted heteroC₉₋₅₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₄₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₃₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₂₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₉alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₈alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₇alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₆alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₄alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₃alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₂alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₁alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₀alkenyl.

In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₅₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₄₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₃₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₂₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₉alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₇alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₆alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₄alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₃₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₂alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₁alkenyl.

In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₅₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₄₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₃₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₂₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₉alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₅₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₇alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₆alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₋₁₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₄alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₃alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₂alkenyl.

In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₅₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₄₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₃₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₂₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₉alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₈alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₇alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₆alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₄alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₃alkenyl.

In some embodiments, R^(L) is optionally substituted heteroC₆alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₇alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₉alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₃alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₄alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₆alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₇alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₅alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₁₉alkenyl. In some embodiments, R^(L) is optionally substituted heteroC₂₀alkenyl.

In some embodiments, for example, in any of the above embodiments, R^(L) is a substituted heteroalkenyl group. In some embodiments, R^(L) is an unsubstituted heteroalkenyl group. In some embodiments, R^(L) is an optionally substituted straight-chain heteroalkenyl group. In some embodiments, R^(L) is a substituted straight-chain heteroalkenyl group. In some embodiments, R^(L) is an unsubstituted straight-chain heteroalkenyl group. In some embodiments, R^(L) is an optionally substituted branched heteroalkenyl group. In some embodiments, R^(L) is a substituted branched heteroalkenyl group. In some embodiments, R^(L) is an unsubstituted branched heteroalkenyl group.

In some embodiments, R^(L) is optionally substituted heteroC₂₋₅₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₄₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₃₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₂₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₉alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₈alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₇alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₆alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₄alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₃alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₂alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₁alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₁₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₉alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₈alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₇alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₋₆alkynyl.

In some embodiments, R^(L) is optionally substituted heteroC₄₋₅₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₄₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₃₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₂₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₉alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₇alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₆alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₄alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₃alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₂alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₁alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₁₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₉alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₇alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₄₋₆alkynyl.

In some embodiments, R^(L) is optionally substituted heteroC₆₋₅₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₄₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₃₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₂₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₉alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₈alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₇alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₆alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₄alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₃alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₂alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₁alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₁₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₉alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₈alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₆₋₇alkynyl.

In some embodiments, R^(L) is optionally substituted heteroC₈₋₅₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₄₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₅₋₃₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₂₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₉alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₅₋₁₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₇alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₆alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₅₋₁₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₄alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₃alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₂alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₁alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₁₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₈₋₉alkynyl.

In some embodiments, R^(L) is optionally substituted heteroC₉₋₅₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₄₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₃₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₂₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₉alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₈alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₇alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₆alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₄alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₃alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₂alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₁alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₉₋₁₀alkynyl.

In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₅₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₄₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₃₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₂₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₉alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₇alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₆alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₄alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₃alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₂alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀₋₁₁alkynyl.

In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₅₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₄₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₋₃₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₂₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₉alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₈alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₇alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₆alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₄alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₃alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁₋₁₂alkynyl.

In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₅₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₄₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₃₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₂₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₉alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₈alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₇alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₆alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₄alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂₋₁₃alkynyl.

In some embodiments, R^(L) is optionally substituted heteroC₆alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₇alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₉alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₀alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₁alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₂alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₃alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₄alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₆alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₇alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₅alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₁₉alkynyl. In some embodiments, R^(L) is optionally substituted heteroC₂₀alkynyl.

In some embodiments, for example, in any of the above embodiments, R^(L) is a substituted heteroalkynyl group. In some embodiments, R^(L) is an unsubstituted heteroalkynyl group. In some embodiments, R^(L) is an optionally substituted straight-chain heteroalkynyl group. In some embodiments, R^(L) is a substituted straight-chain heteroalkynyl group. In some embodiments, R^(L) is an unsubstituted straight-chain heteroalkynyl group. In some embodiments, R^(L) is an optionally substituted branched heteroalkynyl group. In some embodiments, R^(L) is a substituted branched heteroalkynyl group. In some embodiments, R^(L) is an unsubstituted branched heteroalkynyl group.

In some embodiments, R^(L) is a polymer. As used herein, a “polymer”, in some embodiments, refers to a compound comprised of at least 3 (e.g., at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, etc.) repeating covalently bound structural units. The polymer is in certain embodiments biocompatible (i.e., non-toxic). Exemplary polymers include, but are not limited to, cellulose polymers (e.g., hydroxyethylcellulose, ethylcellulose, carboxymethylcellulose, methyl cellulose, hydroxypropylmethylcellulose (HPMC)), dextran polymers, polymaleic acid polymers, poly(acrylic acid) polymers, poly(vinylalcohol) polymers, polyvinylpyrrolidone (PVP) polymers, and polyethyleneglycol (PEG) polymers, and combinations thereof.

In some embodiments, R^(L) is a lipophilic, hydrophobic and/or non-polar group. In some embodiments, R^(L) is a lipophilic group. In some embodiments, R^(L) is a hydrophobic group. In some embodiments, R^(L) is a non-polar group.

In some embodiments, when an R^(L) group is depicted as bisecting a carbon-carbon bond, e.g., of the formula (i), it is understood that R^(L) may be bonded to either carbon.

In some embodiments, at least one instance of R^(Q), R², R⁶, or R⁷ is a group of the formula (i), (ii), or (iii). In some embodiments, at least one instance of R⁶ or R⁷ of R¹ is a group of formula (i), (ii) or (iii). In some embodiments, at least one instance of R⁶ or R⁷ of R¹ is a group of formula (i). In some embodiments, at least one instance of R⁶ or R⁷ of R¹ is a group of formula (i-a). In some embodiments, at least one instance of R⁶ or R⁷ of R¹ is a group of formula (i-a1). In some embodiments, at least one instance of R⁶ or R⁷ of R¹ is a group of formula (i-b). In some embodiments, at least one instance of R⁶ or R⁷ of R¹ is a group of formula (ii). In some embodiments, at least one instance of R⁶ or R⁷ of R¹ is a group of formula (iii).

In some embodiments, the compound (i.e., cationic lipid) of formula I is a compound according to formula II:

or a pharmaceutically acceptable salt thereof,

wherein:

-   -   each m independently is 1 to 19;     -   each n is independently is 1 to 6;     -   each X independently is O or S;     -   each Y independently is O or S;     -   each R_(A) is independently hydrogen, optionally substituted         C1-50 alkyl, optionally substituted C2-50 alkenyl, optionally         substituted C2-50 alkynyl, optionally substituted C3-10         carbocyclyl, optionally substituted 3-14 membered heterocyclyl,         optionally substituted C6-14 aryl, optionally substituted 5-14         membered heteroaryl or halogen, wherein each alkyl, alkenyl,         alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is         independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups,         and

each R_(B) is independently hydrogen, optionally substituted C1-50 alkyl, optionally substituted C2-50 alkenyl, optionally substituted C2-50 alkynyl, optionally substituted C3-10 carbocyclyl, optionally substituted 3-14 membered heterocyclyl, optionally substituted C6-14 aryl, optionally substituted 5-14 membered heteroaryl or halogen, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups.

In some embodiments, each n independently is 2 to 6. In some embodiments, each n independently is 3 to 5. In some embodiments, each n independently is 3 or 4. In some embodiments, each n is the same. In some embodiments, each n is 2. In some embodiments, each n is 3. In some embodiments, each n is 4. In some embodiments, each n is 5. In some embodiments, each n is 6.

In some embodiments, each m independently is 1 to 17. In some embodiments, each m independently is 3 to 15. In some embodiments, each m independently is 5 to 13. In some embodiments, each m independently is 7 to 11. In some embodiments, each m independently is 8 to 10. In some embodiments, each m is the same. In some embodiments, each m is 7. In some embodiments, each m is 8. In some embodiments, each m is 9. In some embodiments, each m is 10. In some embodiments, each m is 11.

In some embodiments, each X is the same. In some embodiments, each X is O. In some embodiments, each X is S. In some embodiments, one X is O and one X is S.

In some embodiments, each Y is the same. In some embodiments, each Y is O. In some embodiments, each Y is S. In some embodiments, one Y is O and one Y is S.

In some embodiments, each X is the same and each Y is the same. In some embodiments, each X is O and each Y is O. In some embodiments, each X is S and each Y is S.

In some embodiments, R_(A) is hydrogen. In some embodiments, R_(A) is optionally substituted C2-50 alkenyl. In some embodiments, R_(A) is optionally substituted C2-50 alkynyl. In some embodiments, R_(A) is optionally substituted C3-10 carbocyclyl. In some embodiments, R_(A) is optionally substituted 3-14 membered heterocyclyl. In some embodiments, R_(A) is optionally substituted C6-14 aryl. In some embodiments, R_(A) is optionally substituted 5-14 membered heteroaryl. In some embodiments, R_(A) is halogen.

In certain embodiments, R_(A) is optionally substituted alkyl; e.g., optionally substituted C₁₋₆alkyl, optionally substituted C₂₋₆alkyl, optionally substituted C₃₋₆alkyl, optionally substituted C₄₋₆alkyl, optionally substituted C₄₋₅alkyl, or optionally substituted C₃₋₄alkyl. In certain embodiments, at least one instance of R_(A) is optionally substituted alkyl; e.g., optionally substituted C₁₋₆alkyl, optionally substituted C₂₋₆alkyl, optionally substituted C₃₋₆alkyl, optionally substituted C₄₋₆alkyl, optionally substituted C₄₋₅alkyl, or optionally substituted C₃₋₄alkyl. In certain embodiments, R_(A) is methyl.

In certain embodiments, R_(A) is optionally substituted alkenyl, e.g., optionally substituted C₂₋₆alkenyl, optionally substituted C₃₋₆alkenyl, optionally substituted C₄₋₆alkenyl, optionally substituted C₄₋₅alkenyl, or optionally substituted C₃₋₄alkenyl. In certain embodiments, at least one instance of R_(A) is optionally substituted alkenyl, e.g., optionally substituted C₂₋₆alkenyl, optionally substituted C₃₋₆alkenyl, optionally substituted C₄₋₆alkenyl, optionally substituted C₄₋₅alkenyl, or optionally substituted C₃₋₄alkenyl.

In certain embodiments, R_(A) is optionally substituted alkynyl, e.g., optionally substituted C₂₋₆alkynyl, optionally substituted C₃₋₆alkynyl, optionally substituted C₄₋₆alkynyl, optionally substituted C₄₋₅alkynyl, or optionally substituted C₃₋₄alkynyl. In certain embodiments, at least one instance of R_(A) is optionally substituted alkynyl, e.g., optionally substituted C₂₋₆alkynyl, optionally substituted C₃₋₆alkynyl, optionally substituted C₄₋₆alkynyl, optionally substituted C₄₋₅alkynyl, or optionally substituted C₃₋₄alkynyl.

In certain embodiments, R_(A) is optionally substituted carbocyclyl, e.g., optionally substituted C₃₋₁₀ carbocyclyl, optionally substituted C₅₋₈ carbocyclyl, optionally substituted C₅₋₆ carbocyclyl, optionally substituted C₅ carbocyclyl, or optionally substituted C₆ carbocyclyl. In certain embodiments, at least one instance of R_(A) is optionally substituted carbocyclyl, e.g., optionally substituted C₃₋₁₀ carbocyclyl, optionally substituted C₅₋₈ carbocyclyl, optionally substituted C₅₋₆ carbocyclyl, optionally substituted C₅ carbocyclyl, or optionally substituted C₆ carbocyclyl.

In some embodiments, R_(A) is optionally substituted heterocyclyl, e.g., optionally substituted 3-14 membered heterocyclyl, optionally substituted 3-10 membered heterocyclyl, optionally substituted 5-8 membered heterocyclyl, optionally substituted 5-6 membered heterocyclyl, optionally substituted 5-membered heterocyclyl, or optionally substituted 6-membered heterocyclyl. In certain embodiments, at least one instance of R_(A) is optionally substituted heterocyclyl, e.g., optionally substituted 3-14 membered heterocyclyl, optionally substituted 3-10 membered heterocyclyl, optionally substituted 5-8 membered heterocyclyl, optionally substituted 5-6 membered heterocyclyl, optionally substituted 5-membered heterocyclyl, or optionally substituted 6-membered heterocyclyl.

In some embodiments, R_(A) is optionally substituted aryl. In some embodiments, R_(A) is optionally substituted phenyl. In some embodiments, R_(A) is phenyl. In some embodiments, R_(A) is substituted phenyl. In certain embodiments, at least one instance of R_(A) is optionally substituted aryl, e.g., optionally substituted phenyl.

In some embodiments, R_(A) is optionally substituted heteroaryl, e.g., optionally substituted 5-14 membered heteroaryl, optionally substituted 5-10 membered heteroaryl, optionally substituted 5-6 membered heteroaryl, optionally substituted 5 membered heteroaryl, or optionally substituted 6 membered heteroaryl. In certain embodiments, at least one instance of R_(A) is optionally substituted heteroaryl, e.g., optionally substituted 5-14 membered heteroaryl, optionally substituted 5-10 membered heteroaryl, optionally substituted 5-6 membered heteroaryl, optionally substituted 5 membered heteroaryl, or optionally substituted 6 membered heteroaryl.

In some embodiments, R_(A) is halogen. In some embodiments, R_(A) is —F. In some embodiments, R_(A) is —Cl. In some embodiments, R_(A) is —Br. In some embodiments, R_(A) is —I.

In some embodiments, R_(B) is hydrogen. In some embodiments, R_(B) is optionally substituted C1-50 alkyl. In some embodiments, R_(B) is optionally substituted C2-50 alkenyl. In some embodiments, R_(B) is optionally substituted C2-50 alkynyl. In some embodiments, R_(B) is optionally substituted C3-10 carbocyclyl. In some embodiments, R_(B) is optionally substituted 3-14 membered heterocyclyl. In some embodiments, R_(B) is optionally substituted C6-14 aryl. In some embodiments, R_(B) is optionally substituted 5-14 membered heteroaryl. In some embodiments, R_(B) is halogen.

In certain embodiments, R_(B) is optionally substituted alkyl; e.g., optionally substituted C₁₋₆alkyl, optionally substituted C₂₋₆alkyl, optionally substituted C₃₋₆alkyl, optionally substituted C₄₋₆alkyl, optionally substituted C₄₋₅alkyl, or optionally substituted C₃₋₄alkyl. In certain embodiments, at least one instance of R_(B) is optionally substituted alkyl; e.g., optionally substituted C₁₋₆alkyl, optionally substituted C₂₋₆alkyl, optionally substituted C₃₋₆alkyl, optionally substituted C₄₋₆alkyl, optionally substituted C₄₋₅alkyl, or optionally substituted C₃₋₄alkyl. In certain embodiments, R_(B) is methyl.

In certain embodiments, R_(B) is optionally substituted alkenyl, e.g., optionally substituted C₂₋₆alkenyl, optionally substituted C₃₋₆alkenyl, optionally substituted C₄₋₆alkenyl, optionally substituted C₄₋₅alkenyl, or optionally substituted C₃₋₄alkenyl. In certain embodiments, at least one instance of R_(B) is optionally substituted alkenyl, e.g., optionally substituted C₂₋₆alkenyl, optionally substituted C₃₋₆alkenyl, optionally substituted C₄₋₆alkenyl, optionally substituted C₄₋₅alkenyl, or optionally substituted C₃₋₄alkenyl.

In certain embodiments, R_(B) is optionally substituted alkynyl, e.g., optionally substituted C₂₋₆alkynyl, optionally substituted C₃₋₆alkynyl, optionally substituted C₄₋₆alkynyl, optionally substituted C₄₋₅alkynyl, or optionally substituted C₃₋₄alkynyl. In certain embodiments, at least one instance of R_(B) is optionally substituted alkynyl, e.g., optionally substituted C₂₋₆alkynyl, optionally substituted C₃₋₆alkynyl, optionally substituted C₄₋₆alkynyl, optionally substituted C₄₋₅alkynyl, or optionally substituted C₃₋₄alkynyl.

In certain embodiments, R_(B) is optionally substituted carbocyclyl, e.g., optionally substituted C₃₋₁₀ carbocyclyl, optionally substituted C₅₋₈ carbocyclyl, optionally substituted C₅₋₆ carbocyclyl, optionally substituted C₅ carbocyclyl, or optionally substituted C₆ carbocyclyl. In certain embodiments, at least one instance of R_(B) is optionally substituted carbocyclyl, e.g., optionally substituted C₃₋₁₀ carbocyclyl, optionally substituted C₅₋₈ carbocyclyl, optionally substituted C₅₋₆ carbocyclyl, optionally substituted C₅ carbocyclyl, or optionally substituted C₆ carbocyclyl.

In some embodiments, R_(B) is optionally substituted heterocyclyl, e.g., optionally substituted 3-14 membered heterocyclyl, optionally substituted 3-10 membered heterocyclyl, optionally substituted 5-8 membered heterocyclyl, optionally substituted 5-6 membered heterocyclyl, optionally substituted 5-membered heterocyclyl, or optionally substituted 6-membered heterocyclyl. In certain embodiments, at least one instance of R_(B) is optionally substituted heterocyclyl, e.g., optionally substituted 3-14 membered heterocyclyl, optionally substituted 3-10 membered heterocyclyl, optionally substituted 5-8 membered heterocyclyl, optionally substituted 5-6 membered heterocyclyl, optionally substituted 5-membered heterocyclyl, or optionally substituted 6-membered heterocyclyl.

In some embodiments, R_(B) is optionally substituted aryl. In some embodiments, R_(B) is optionally substituted phenyl. In some embodiments, R_(B) is phenyl. In some embodiments, R_(B) is substituted phenyl. In certain embodiments, at least one instance of R_(B) is optionally substituted aryl, e.g., optionally substituted phenyl.

In some embodiments, R_(B) is optionally substituted heteroaryl, e.g., optionally substituted 5-14 membered heteroaryl, optionally substituted 5-10 membered heteroaryl, optionally substituted 5-6 membered heteroaryl, optionally substituted 5 membered heteroaryl, or optionally substituted 6 membered heteroaryl. In certain embodiments, at least one instance of R_(B) is optionally substituted heteroaryl, e.g., optionally substituted 5-14 membered heteroaryl, optionally substituted 5-10 membered heteroaryl, optionally substituted 5-6 membered heteroaryl, optionally substituted 5 membered heteroaryl, or optionally substituted 6 membered heteroaryl.

In some embodiments, R_(B) is halogen. In some embodiments, R_(B) is —F. In some embodiments, R_(B) is —Cl. In some embodiments, R_(B) is —Br. In some embodiments, R_(B) is —I.

In some embodiments, at least one R_(A) is optionally substituted C1-50 alkyl. In some embodiments, at least one R_(A) and one R_(B) are optionally substituted C1-50 alkyl. In some embodiments, at least one R_(A) and one R_(B) are optionally substituted C1-50 alkyl, where the R_(A) and R_(B) are attached to the same carbon. In some embodiments, at least one R_(A) is methyl. In some embodiments, at least one R_(A) and one R_(B) are methyl. In some embodiments, at least one R_(A) and one R_(B) are methyl, where the R_(A) and R_(B) are attached to the same carbon.

In some embodiments, a compound of formula II is a compound of formula II-a:

or a pharmaceutically acceptable salt thereof, wherein m and n are as defined above and described herein.

In some embodiments, a compound of formula II is a compound of formula II-a wherein each n independently is 2 to 6 and each m independently is 3 to 15. In some embodiments, a compound of formula II is a compound of formula II-a wherein each n independently is 3 to 5 and each m independently is 5 to 13. In some embodiments, a compound of formula II is a compound of formula II-a wherein each n independently is 3 or 4 and each m independently is 7 to 11. In some embodiments, a compound of formula II is a compound of formula II-a wherein each n is the same and is 3 or 4, and each m is the same and is 7 to 11.

In some embodiments, a compound of formula II is a compound of formula III:

or a pharmaceutically acceptable salt thereof.

In some embodiments, a compound of formula II is a compound of formula IV:

or a pharmaceutically acceptable salt thereof.

In some embodiments, a compound of formula II is a compound of formula V:

or a pharmaceutically acceptable salt thereof. Liposomes for the Delivery of Agents, Such as mRNA

Among other things, the present invention provides composition comprising a biodegradable compound described herein for delivery of therapeutic agents. In some embodiments, a composition provided is a lipid based nanoparticle, such as a liposome. As used herein, the term “liposome” refers to any lamellar, multilamellar, or solid lipid nanoparticle vesicle. Typically, a liposome as used herein can be formed by mixing one or more lipids or by mixing one or more lipids and polymer(s). Thus, the term “liposome” as used herein encompasses both lipid and polymer based nanoparticles. In particular, a liposome according to the present invention incorporates a biodegradable compound described herein as a cationic lipid. As a non-limiting example, a liposome according to the present invention is a compound of formula III i.e., 3,6-bis(4-(bis(2-hydroxydodecyl)amino)butyl)-1,4-dioxane-2,5-dione. A suitable liposome may also contain second or additional cationic lipids, helper lipids (e.g., non-cationic lipids and/or cholesterol-based lipids), PEG-modified lipids, and/or polymers.

In some embodiments, cationic lipid(s) (e.g., the compound of formula III) constitute(s) about 30-50% (e.g., about 30-45%, about 30-40%, about 35-50%, about 35-45%, or about 35-40%) of the liposome by molar ratio. In some embodiments, the cationic lipid (e.g., the compound of formula III) constitutes about 30%, about 35%, about 40%, about 45%, or about 50% of the liposome by molar ratio. In some embodiments, the liposome comprises a second lipid or additional cationic lipids.

Second or Additional Cationic Lipids

In some embodiments, liposomes may comprise a second or additional cationic lipid. As used herein, the phrase “cationic lipid” refers to any of a number of lipid species that have a net positive charge at a selected pH, such as physiological pH. Several cationic lipids have been described in the literature, many of which are commercially available. Particularly suitable cationic lipids for use in the compositions and methods of the invention include those described in international patent publications WO 2010/053572 (and particularly, C12-200 described at paragraph [00225]), WO 2012/170930 and WO 2013063468 each of which is incorporated herein by reference in its entirety. In certain embodiments, the compositions and methods of the invention employ a lipid nanoparticles comprising an ionizable (titratable) cationic lipid described in International Patent Application No. PCT/US2013/034602, filed Mar. 29, 2013, Publ. No. WO 2013/149140 (incorporated herein by reference), such as, e.g, (15Z, 18Z)—N,N-dimethyl-6-(9Z, 12Z)-octadeca-9, 12-dien-1-yl)tetracosa-15,18-dien-1-amine (HGT5000), (15Z, 18Z)—N,N-dimethyl-6-((9Z, 12Z)-octadeca-9, 12-dien-1-yl)tetracosa-4,15,18-trien-1-amine (HGT5001), and (15Z, 18Z)—N,N-dimethyl-6-((9Z, 12Z)-octadeca-9, 12-dien-1-yl)tetracosa-5, 15, 18-trien-1-amine (HGT5002).

In some embodiments, the second or additional cationic lipid is cKK-E12 (3,6-bis(4-(bis(2-hydroxydodecyl)amino)butyl)piperazine-2,5-dione) or derivatives thereof, as described in international patent publications WO 2013/063468 incorporated herein by reference in its entirety.

In some embodiments, the second or additional cationic lipid N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride or “DOTMA” is used. (Feigner et al. (Proc. Nat'l Acad. Sci. 84, 7413 (1987); U.S. Pat. No. 4,897,355). DOTMA can be formulated alone or can be combined with the neutral lipid, dioleoylphosphatidyl-ethanolamine or “DOPE” or other cationic or non-cationic lipids into a liposomal transfer vehicle or a lipid nanoparticle, and such liposomes can be used to enhance the delivery of nucleic acids into target cells. Other suitable cationic lipids include, for example, 5-carboxyspermylglycinedioctadecylamide or “DOGS,” 2,3-dioleyloxy-N-[2(spermine-carboxamido)ethyl]-N,N-dimethyl-1-propanaminium or “DOSPA” (Behr et al. Proc. Nat.'l Acad. Sci. 86, 6982 (1989); U.S. Pat. No. 5,171,678; U.S. Pat. No. 5,334,761), 1,2-Dioleoyl-3-Dimethylammonium-Propane or “DODAP”, 1,2-Dioleoyl-3-Trimethylammonium-Propane or “DOTAP”. Additional exemplary cationic lipids also include 1,2-distearyloxy-N,N-dimethyl-3-aminopropane or “DSDMA”, 1,2-dioleyloxy-N,N-dimethyl-3-aminopropane or “DODMA”, 1,2-dilinoleyloxy-N,N-dimethyl-3-aminopropane or “DLinDMA”, 1,2-dilinolenyloxy-N,N-dimethyl-3-aminopropane or “DLenDMA”, N-dioleyl-N,N-dimethylammonium chloride or “DODAC”, N,N-distearyl-N,N-dimethylarnrnonium bromide or “DDAB”, N-(1,2-dimyristyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethyl ammonium bromide or “DMRIE”, 3-dimethylamino-2-(cholest-5-en-3-beta-oxybutan-4-oxy)-1-(cis,cis-9,12-octadecadienoxy)propane or “CLinDMA”, 2-[5′-(cholest-5-en-3-beta-oxy)-3′-oxapentoxy)-3-dimethy 1-1-(cis,cis-9′, 1-2′-octadecadienoxy)propane or “CpLinDMA”, N,N-dimethyl-3,4-dioleyloxybenzylamine or “DMOBA”, 1,2-N,N′-dioleylcarbamyl-3-dimethylaminopropane or “DOcarbDAP”, 2,3-Dilinoleoyloxy-N,N-dimethylpropylamine or “DLinDAP”, 1,2-N,N′-Dilinoleylcarbamyl-3-dimethylaminopropane or “DLincarbDAP”, 1,2-Dilinoleoylcarbamyl-3-dimethylaminopropane or “DLinCDAP”, 2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane or “DLin--DMA”, 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane or “DLin-K-XTC2-DMA”, and 2-(2,2-di((9Z, 12Z)-octadeca-9,12-dien-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylethanamine (DLin-KC2-DMA)) (See, WO 2010/042877; Semple et al., Nature Biotech. 28: 172-176 (2010)), or mixtures thereof. (Heyes, J., et al., J Controlled Release 107: 276-287 (2005); Morrissey, D V., et al., Nat. Biotechnol. 23(8): 1003-1007 (2005); PCT Publication WO2005/121348A1). In some embodiments, one or more of the cationic lipids comprise at least one of an imidazole, dialkylamino, or guanidinium moiety.

In some embodiments, the second or additional cationic lipid may be chosen from XTC (2,2-Dilinoleyl 1-4-dimethylaminoethy 1-[1,3]-dioxolane), MC3 (((6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate), ALNY-100 ((3aR,5s,6aS)—N,N-dimethyl-2,2-di((9Z,12Z)-octadeca-9,12-dienyl)tetrahydro-3aH-cyclopenta[d][1,3]dioxol-5-amine)), NC98-5 (4,7,13-tris(3-oxo-3-(undecylamino)propyl)-N1,N16-diundecyl-4,7,10,13-tetraazahexadecane-1,16-diamide), DODAP (1,2-dioleyl-3-dimethylammonium propane), HGT4003 (WO 2012/170889, the teachings of which are incorporated herein by reference in their entirety), ICE (WO 2011/068810, the teachings of which are incorporated herein by reference in their entirety), HGT5000 (international patent publication WO/2013/149140, the teachings of which are incorporated herein by reference in their entirety) or HGT5001 (cis or trans) (WO/2013/149140), aminoalcohol lipidoids such as those disclosed in WO2010/053572, DOTAP (1,2-dioleyl-3-trimethylammonium propane), DOTMA (1,2-di-O-octadecenyl-3-trimethylammonium propane), DLinDMA (Heyes, J.; Palmer, L.; Bremner, K.; MacLachlan, I. “Cationic lipid saturation influences intracellular delivery of encapsulated nucleic acids” J. Contr. Rel. 2005, 107, 276-287), DLin-KC2-DMA (Semple, S. C. et al. “Rational Design of Cationic Lipids for siRNA Delivery” Nature Biotech. 2010, 28, 172-176), C12-200 (Love, K. T. et al. “Lipid-like materials for low-dose in vivo gene silencing” PNAS 2010, 107, 1864-1869).

Non-cationic/Helper Lipids

In some embodiments, provided liposomes contain one or more non-cationic (“helper”) lipids. As used herein, the phrase “non-cationic lipid” refers to any neutral, zwitterionic or anionic lipid. As used herein, the phrase “anionic lipid” refers to any of a number of lipid species that carry a net negative charge at a selected H, such as physiological pH. Non-cationic lipids include, but are not limited to, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, 1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), or a mixture thereof.

In some embodiments, such non-cationic lipids may be used alone, but are preferably used in combination with other excipients, for example, cationic lipids. In some embodiments, the non-cationic lipid may comprise a molar ratio of about 5% to about 90%, or about 10% to about 70% of the total lipid present in a liposome. In some embodiments, a non-cationic lipid is a neutral lipid, i.e., a lipid that does not carry a net charge in the conditions under which the composition is formulated and/or administered. In some embodiments, the percentage of non-cationic lipid in a liposome may be greater than 5%, greater than 10%, greater than 20%, greater than 30%, or greater than 40%.

Cholesterol-Based Lipids

In some embodiments, provided liposomes comprise one or more cholesterol-based lipids. For example, suitable cholesterol-based cationic lipids include, for example, DC-Chol (N,N-dimethyl-N-ethylcarboxamidocholesterol), 1,4-bis(3-N-oleylamino-propyl)piperazine (Gao, et al. Biochem. Biophys. Res. Comm. 179, 280 (1991); Wolf et al. BioTechniques 23, 139 (1997); U.S. Pat. No. 5,744,335), or ICE. In some embodiments, the cholesterol-based lipid may comprise a molar ration of about 2% to about 30%, or about 5% to about 20% of the total lipid present in a liposome. In some embodiments, The percentage of cholesterol-based lipid in the lipid nanoparticle may be greater than 5, %, 10%, greater than 20%, greater than 30%, or greater than 40%.

PEGylated Lipids

In some embodiments, provided liposomes comprise one or more PEGylated lipids. For example, the use of polyethylene glycol (PEG)-modified phospholipids and derivatized lipids such as derivatized ceramides (PEG-CER), including N-Octanoyl-Sphingosine-1-[Succinyl(Methoxy Polyethylene Glycol)-2000] (C8 PEG-2000 ceramide) is also contemplated by the present invention in combination with one or more of the cationic and, in some embodiments, other lipids together which comprise the liposome. Contemplated PEG-modified lipids include, but are not limited to, a polyethylene glycol chain of up to 5 kDa in length covalently attached to a lipid with alkyl chain(s) of C₆-C₂₀ length. In some embodiments, a PEG-modified or PEGylated lipid is PEGylated cholesterol or PEG-2K. The addition of such components may prevent complex aggregation and may also provide a means for increasing circulation lifetime and increasing the delivery of the lipid-nucleic acid composition to the target cell, (Klibanov et al. (1990) FEBS Letters, 268 (1): 235-237), or they may be selected to rapidly exchange out of the formulation in vivo (see U.S. Pat. No. 5,885,613).

In some embodiments, particularly useful exchangeable lipids are PEG-ceramides having shorter acyl chains (e.g., C₁₄ or C₁₈). The PEG-modified phospholipid and derivitized lipids of the present invention may comprise a molar ratio from about 0% to about 15%, about 0.5% to about 15%, about 1% to about 15%, about 4% to about 10%, or about 2% of the total lipid present in the liposome.

According to various embodiments, the selection of second or additional cationic lipids, non-cationic lipids and/or PEG-modified lipids which comprise the lipid nanoparticle, as well as the relative molar ratio of such lipids to each other, is based upon the characteristics of the selected lipid(s), the nature of the intended target cells, the characteristics of the mRNA to be delivered. Additional considerations include, for example, the saturation of the alkyl chain, as well as the size, charge, pH, pKa, fusogenicity and toxicity of the selected lipid(s). Thus the molar ratios may be adjusted accordingly. In some embodiments, the percentage of PEG-modified lipid in a liposome may be greater than 1%, greater than 2%, greater than 5%, greater than 10%, or greater than 15%.

Polymers

In some embodiments, a suitable liposome according to the present invention further includes a polymer, in combination with one or more cationic lipids as described and, in some embodiments, other carriers including various lipids described herein. Thus, in some embodiments, liposomal delivery vehicles, as used herein, also encompass polymer containing nanoparticles. Suitable polymers may include, for example, polyacrylates, polyalkycyanoacrylates, polylactide, polylactide-polyglycolide copolymers, polycaprolactones, dextran, albumin, gelatin, alginate, collagen, chitosan, cyclodextrins, protamine, PEGylated protamine, PLL, PEGylated PLL and polyethylenimine (PEI). When PEI is present, it may be branched PEI of a molecular weight ranging from 10 to 40 kDA, e.g., 25 kDa branched PEI (Sigma #408727).

Therapeutic Agents

The present invention may be used to delivery any therapeutic agents. Specifically, any therapeutic agents to be administered to a subject may be delivered using the complexes, picoparticles, nanoparticles, microparticles, micelles, or liposomes, described herein. The agent may be an organic molecule (e.g., a therapeutic agent, a drug), inorganic molecule, nucleic acid, protein, amino acid, peptide, polypeptide, polynucleotide, targeting agent, isotopically labeled organic or inorganic molecule, vaccine, immunological agent, etc. In certain embodiments of the present invention, the agent to be delivered may be a mixture of agents.

In certain embodiments, the therapeutic agents are organic molecules with pharmaceutical activity, e.g., a drug. In certain embodiments, the drug is an antibiotic, anti-viral agent, anesthetic, steroidal agent, anti-inflammatory agent, anti-neoplastic agent, anti-cancer agent, antigen, vaccine, antibody, decongestant, antihypertensive, sedative, birth control agent, progestational agent, anti-cholinergic, analgesic, anti-depressant, anti-psychotic, I3-adrenergic blocking agent, diuretic, cardiovascular active agent, vasoactive agent, non-steroidal anti-inflammatory agent, nutritional agent, etc.

Diagnostic agents include gases; metals; commercially available imaging agents used in positron emissions tomography (PET), computer assisted tomography (CAT), single photon emission computerized tomography, x-ray, fluoroscopy, and magnetic resonance imaging (MRI); and contrast agents. Examples of suitable materials for use as contrast agents in MRI include gadolinium chelates, as well as iron, magnesium, manganese, copper, and chromium. Examples of materials useful for CAT and x-ray imaging include iodinebased materials.

Therapeutic and prophylactic agents include, but are not limited to, antibiotics, nutritional supplements, and vaccines. Vaccines may comprise isolated proteins or peptides, inactivated organisms and viruses, dead organisms and viruses, genetically altered organisms or viruses, and cell extracts.

Polynucleotides

The present invention may be used to deliver any polynucleotide. In certain embodiments, the polynucleotide is an interfering RNA (RNAi). The phenomenon of RNAi is discussed in greater detail, for example, in the following references: Elbashir et al., 2001, Genes Dev., 15:188; Fire et al., 1998, Nature, 391:806; Tabara et al., 1999, Cell, 99:123; Hammond et al., Nature, 2000, 404:293; Zamore et al., 2000, Cell, 101:25; Chakraborty, 2007, Curr. Drug Targets, 8:469; and Morris and Rossi, 2006, Gene Ther., 13:553. In certain embodiments, the polynucleotide is a dsRNA (double-stranded RNA). In certain embodiments, the polynucleotide is an siRNA (short interfering RNA). In certain embodiments, the polynucleotide is an shRNA (short hairpin RNA). In certain embodiments, the polynucleotide is an miRNA (micro RNA). Micro RNAs (miRNAs) are genomically encoded non-coding RNAs of about 21-23 nucleotides in length that help regulate gene expression, particularly during development. See, e.g., Bartel, 2004, Cell, 116:281; Novina and Sharp, 2004, Nature, 430:161; and U.S. Patent Publication 2005/0059005; also reviewed in Wang and Li, 2007, Front. Biosci., 12:3975; and Zhao, 2007, Trends Biochem. Sci., 32:189. In certain embodiments, the polynucleotide is an antisense RNA.

In certain embodiments, the polynucleotide may be provided as an antisense agent or RNA interference (RNAi). See, e.g., Fire et al., Nature 391:806-811, 1998. Antisense therapy is meant to include, e.g., administration or in situ provision of single- or double-stranded oligonucleotides or their derivatives which specifically hybridize, e.g., bind, under cellular conditions, with cellular mRNA and/or genomic DNA, or mutants thereof, so as to inhibit expression of the encoded protein, e.g., by inhibiting transcription and/or translation. See, e.g., Crooke “Molecular mechanisms of action of antisense drugs” Biochim. Biophys. Acta 1489(1):31-44, 1999; Crooke “Evaluating the mechanism of action of antiproliferative antisense drugs” Antisense Nucleic Acid Drug Dev. 10(2):123-126, discussion 127, 2000; Methods in Enzymology volumes 313-314, 1999. The binding may be by conventional base pair complementarity, or, for example, in the case of binding to DNA duplexes, through specific interactions in the major groove of the double helix (i.e., triple helix formation). See, e.g., Chan et al., J. Mol. Med. 75(4):267-282, 1997.

In some embodiments, dsRNA, siRNA, shRNA, miRNA, antisense RNA, and/or RNAi can be designed and/or predicted using one or more of a large number of available algorithms. To give but a few examples, the following resources can be utilized to design and/or predict polynucleotides: algorithms found at Alnylum Online, Dharmacon Online, OligoEngine Online, Molecula Online, Ambion Online, BioPredsi Online, RNAi Web Online, Chang Bioscience Online, Invitrogen Online, LentiWeb Online GenScript Online, Protocol Online; Reynolds et al., 2004, Nat. Biotechnol., 22:326; Naito et al., 2006, Nucleic Acids Res., 34:W448; Li et al., 2007, RNA, 13:1765; Yiu et al., 2005, Bioinformatics, 21:144; and Jia et al., 2006, BMC Bioinformatics, 7: 271.

The polynucleotides may be of any size or sequence, and they may be single- or double-stranded. In certain embodiments, the polynucleotide is greater than 100 base pairs long. In certain embodiments, the polynucleotide is greater than 1000 base pairs long and may be greater than 10,000 base pairs long. The polynucleotide may be provided by any means known in the art. In certain embodiments, the polynucleotide has been engineered using recombinant techniques. See, e.g., Ausubel et al., Current Protocols in Molecular Biology (John Wiley & Sons, Inc., New York, 1999); Molecular Cloning: A Laboratory Manual, 2nd Ed., ed. by Sambrook, Fritsch, and Maniatis (Cold Spring Harbor Laboratory Press: 1989). The polynucleotide may also be obtained from natural sources and purified from contaminating components found normally in nature. The polynucleotide may also be chemically synthesized in a laboratory. In certain embodiments, the polynucleotide is synthesized using standard solid phase chemistry.

The polynucleotide may be modified by chemical or biological means. In certain embodiments, these modifications lead to increased stability of the polynucleotide. Modifications include methylation, phosphorylation, end-capping, etc.

mRNA

The present invention can be used to deliver any mRNA. mRNA is typically thought of as the type of RNA that carries information from DNA to the ribosome. The existence of mRNA is usually very brief and includes processing and translation, followed by degradation. Typically, in eukaryotic organisms, mRNA processing comprises the addition of a “cap” on the N-terminal (5′) end, and a “tail” on the C-terminal (3′) end. A typical cap is a 7-methylguanosine cap, which is a guanosine that is linked through a 5′-5′-triphosphate bond to the first transcribed nucleotide. The presence of the cap is important in providing resistance to nucleases found in most eukaryotic cells. The tail is typically a polyadenylation event whereby a polyadenylyl moiety is added to the 3′ end of the mRNA molecule. The presence of this “tail” serves to protect the mRNA from exonuclease degradation. Messenger RNA typically is translated by the ribosomes into a series of amino acids that make up a protein.

Any mRNA capable of being translated into one or more peptides (e.g., proteins) or peptide fragments is contemplated as within the scope of the present invention. In some embodiments, an mRNA encodes one or more naturally occurring peptides. In some embodiments, an mRNA encodes one or more modified or non-natural peptides.

In some embodiments an mRNA encodes an intracellular protein. In some embodiments, an mRNA encodes a cytosolic protein. In some embodiments, an mRNA encodes a protein associated with the actin cytoskeleton. In some embodiments, an mRNA encodes a protein associated with the plasma membrane. In some specific embodiments, an mRNA encodes a transmembrane protein. In some specific embodiments an mRNA encodes an ion channel protein. In some embodiments, an mRNA encodes a perinuclear protein. In some embodiments, an mRNA encodes a nuclear protein. In some specific embodiments, an mRNA encodes a transcription factor. In some embodiments, an mRNA encodes a chaperone protein. In some embodiments, an mRNA encodes an intracellular enzyme (e.g., mRNA encoding an enzyme associated with urea cycle or lysosomal storage metabolic disorders). In some embodiments, an mRNA encodes a protein involved in cellular metabolism, DNA repair, transcription and/or translation. In some embodiments, an mRNA encodes an extracellular protein. In some embodiments, an mRNA encodes a protein associated with the extracellular matrix. In some embodiments an mRNA encodes a secreted protein. In specific embodiments, an mRNA used in the composition and methods of the invention may be used to express functional proteins or enzymes that are excreted or secreted by one or more target cells into the surrounding extracellular fluid (e.g., mRNA encoding hormones and/or neurotransmitters).

Synthesis of mRNA

mRNAs according to the present invention may be synthesized according to any of a variety of known methods. For example, mRNAs according to the present invention may be synthesized via in vitro transcription (IVT). Briefly, IVT is typically performed with a linear or circular DNA template containing a promoter, a pool of ribonucleotide triphosphates, a buffer system that may include DTT and magnesium ions, and an appropriate RNA polymerase (e.g., T3, T7 or SP6 RNA polymerase), DNAse I, pyrophosphatase, and/or RNAse inhibitor. The exact conditions will vary according to the specific application.

In some embodiments, for the preparation of mRNA according to the invention, a DNA template is transcribed in vitro. A suitable DNA template typically has a promoter, for example a T3, T7 or SP6 promoter, for in vitro transcription, followed by desired nucleotide sequence for desired mRNA and a termination signal.

Desired mRNA sequence(s) according to the invention may be determined and incorporated into a DNA template using standard methods. For example, starting from a desired amino acid sequence (e.g., an enzyme sequence), a virtual reverse translation is carried out based on the degenerated genetic code. Optimization algorithms may then be used for selection of suitable codons. Typically, the G/C content can be optimized to achieve the highest possible G/C content on one hand, taking into the best possible account the frequency of the tRNAs according to codon usage on the other hand. The optimized RNA sequence can be established and displayed, for example, with the aid of an appropriate display device and compared with the original (wild-type) sequence. A secondary structure can also be analyzed to calculate stabilizing and destabilizing properties or, respectively, regions of the RNA.

Modified mRNA

In some embodiments, mRNA according to the present invention may be synthesized as unmodified or modified mRNA. Typically, mRNAs are modified to enhance stability. Modifications of mRNA can include, for example, modifications of the nucleotides of the RNA. An modified mRNA according to the invention can thus include, for example, backbone modifications, sugar modifications or base modifications. In some embodiments, mRNAs may be synthesized from naturally occurring nucleotides and/or nucleotide analogues (modified nucleotides) including, but not limited to, purines (adenine (A), guanine (G)) or pyrimidines (thymine (T), cytosine (C), uracil (U)), and as modified nucleotides analogues or derivatives of purines and pyrimidines, such as e.g. 1-methyl-adenine, 2-methyl-adenine, 2-methylthio-N-6-isopentenyl-adenine, N6-methyl-adenine, N6-isopentenyl-adenine, 2-thio-cytosine, 3-methyl-cytosine, 4-acetyl-cytosine, 5-methyl-cytosine, 2,6-diaminopurine, 1-methyl-guanine, 2-methyl-guanine, 2,2-dimethyl-guanine, 7-methyl-guanine, inosine, 1-methyl-inosine, pseudouracil (5-uracil), dihydro-uracil, 2-thio-uracil, 4-thio-uracil, 5-carboxymethylamino-methyl-2-thio-uracil, 5-(carboxyhydroxymethyl)-uracil, 5-fluoro-uracil, 5-bromo-uracil, 5-carboxymethylaminomethyl-uracil, 5-methyl-2-thio-uracil, 5-methyl-uracil, N-uracil-5-oxy-acetic acid methyl ester, 5-methylaminomethyl-uracil, 5-methoxyaminomethyl-2-thio-uracil, 5′-methoxycarbonylmethyl-uracil, 5-methoxy-uracil, uracil-5-oxyacetic acid methyl ester, uracil-5-oxyacetic acid (v), 1-methyl-pseudouracil, queosine, 3-D-mannosyl-queosine, wybutosine, and phosphoramidates, phosphorothioates, peptide nucleotides, methylphosphonates, 7-deaza-guanosine, 5-methylcytosine and inosine. The preparation of such analogues is known to a person skilled in the art e.g. from the U.S. Pat. No. 4,373,071, U.S. Pat. No. 4,401,796, U.S. Pat. No. 4,415,732, U.S. Pat. No. 4,458,066, U.S. Pat. No. 4,500,707, U.S. Pat. No. 4,668,777, U.S. Pat. No. 4,973,679, U.S. Pat. No. 5,047,524, U.S. Pat. No. 5,132,418, U.S. Pat. No. 5,153,319, U.S. Pat. Nos. 5,262,530 and 5,700,642, the disclosures of which are incorporated by reference in their entirety.

In some embodiments, mRNAs (e.g., enzyme encoding mRNAs) may contain RNA backbone modifications. Typically, a backbone modification is a modification in which the phosphates of the backbone of the nucleotides contained in the RNA are modified chemically. Exemplary backbone modifications typically include, but are not limited to, modifications from the group consisting of methylphosphonates, methylphosphoramidates, phosphoramidates, phosphorothioates (e.g. cytidine 5′-O-(1-thiophosphate)), boranophosphates, positively charged guanidinium groups etc., which means by replacing the phosphodiester linkage by other anionic, cationic or neutral groups.

In some embodiments, mRNAs (e.g., enzyme encoding mRNAs) may contain sugar modifications. A typical sugar modification is a chemical modification of the sugar of the nucleotides it contains including, but not limited to, sugar modifications chosen from the group consisting of 2′-deoxy-2′-fluoro-oligoribonucleotide (2′-fluoro-2′-deoxycytidine 5′-triphosphate, 2′-fluoro-2′-deoxyuridine 5′-triphosphate), 2′-deoxy-2′-deamine-oligoribonucleotide (2′-amino-2′-deoxycytidine 5′-triphosphate, 2′-amino-2′-deoxyuridine 5′-triphosphate), 2′-O-alkyloligoribo-nucleotide, 2′-deoxy-2′-C-alkyloligoribonucleotide (2′-O-methylcytidine 5′-triphosphate, 2′-methyluridine 5′-triphosphate), 2′-C-alkyloligoribonucleotide, and isomers thereof (2′-ara-cytidine 5′-triphosphate, 2′-arauridine 5′-triphosphate), or azidotriphosphates (2′-azido-2′-deoxy-cytidine 5′-triphosphate, 2′-azido-2′-deoxyuridine 5′-triphosphate).

In some embodiments, mRNAs (e.g., enzyme encoding mRNAs) may contain modifications of the bases of the nucleotides (base modifications). A modified nucleotide which contains a base modification is also called a base-modified nucleotide. Examples of such base-modified nucleotides include, but are not limited to, 2-amino-6-chloropurine riboside 5′-tri-phosphate, 2-aminoadenosine 5′-triphosphate, 2-thiocytidine 5′-triphosphate, 2-thiouridine 5′-tri-phosphate, 4-thiouridine 5′-triphosphate, 5-aminoallylcytidine 5′-triphosphate, 5-aminoallyl-uridine 5′-triphosphate, 5-bromocytidine 5′-triphosphate, 5-bromouridine 5′-triphosphate, 5-iodo-cytidine 5′-triphosphate, 5-iodouridine 5′-triphosphate, 5-methylcytidine 5′-triphosphate, 5-methyluridine 5′-triphosphate, 6-azacytidine 5′-triphosphate, 6-azauridine 5′-triphosphate, 6-chloropurine riboside 5′-triphosphate, 7-deazaadenosine 5′-triphosphate, 7-deazaguanosine 5′-triphosphate, 8-azaadenosine 5′-triphosphate, 8-azidoadenosine 5′-triphosphate, benzimidazole riboside 5′-triphosphate, N1-methyladenosine 5′-triphosphate, N1-methylguanosine 5′-tri-phosphate, N6-methyladenosine 5′-triphosphate, 06-methylguanosine 5′-triphosphate, pseudo-uridine 5′-triphosphate, puromycin 5′-triphosphate or xanthosine 5′-triphosphate.

Cap Structure

Typically, mRNA synthesis includes the addition of a “cap” on the N-terminal (5′) end, and a “tail” on the C-terminal (3′) end. The presence of the cap is important in providing resistance to nucleases found in most eukaryotic cells. The presence of a “tail” serves to protect the mRNA from exonuclease degradation.

Thus, in some embodiments, mRNAs (e.g., enzyme encoding mRNAs) include a 5′ cap structure. A 5′ cap is typically added as follows: first, an RNA terminal phosphatase removes one of the terminal phosphate groups from the 5′ nucleotide, leaving two terminal phosphates; guanosine triphosphate (GTP) is then added to the terminal phosphates via a guanylyl transferase, producing a 5′5′5 triphosphate linkage; and the 7-nitrogen of guanine is then methylated by a methyltransferase. Examples of cap structures include, but are not limited to, m7G(5′)ppp (5′(A,G(5′)ppp(5′)A and G(5′)ppp(5′)G.

In some embodiments, naturally occurring cap structures comprise a 7-methyl guanosine that is linked via a triphosphate bridge to the 5′-end of the first transcribed nucleotide, resulting in a dinucleotide cap of m⁷G(5′)ppp(5′)N, where N is any nucleoside. In vivo, the cap is added enzymatically. The cap is added in the nucleus and is catalyzed by the enzyme guanylyl transferase. The addition of the cap to the 5′ terminal end of RNA occurs immediately after initiation of transcription. The terminal nucleoside is typically a guanosine, and is in the reverse orientation to all the other nucleotides, i.e., G(5′)ppp(5′)GpNpNp.

A common cap for mRNA produced by in vitro transcription is m⁷G(5′)ppp(5′)G, which has been used as the dinucleotide cap in transcription with T7 or SP6 RNA polymerase in vitro to obtain RNAs having a cap structure in their 5′-termini. The prevailing method for the in vitro synthesis of capped mRNA employs a pre-formed dinucleotide of the form m⁷G(5′)ppp(5′)G (“m⁷GpppG”) as an initiator of transcription.

To date, a usual form of a synthetic dinucleotide cap used in in vitro translation experiments is the Anti-Reverse Cap Analog (“ARCA”) or modified ARCA, which is generally a modified cap analog in which the 2′ or 3′ OH group is replaced with —OCH₃.

Additional cap analogs include, but are not limited to, chemical structures selected from the group consisting of m⁷GpppG, m⁷GpppA, m⁷GpppC; unmethylated cap analogs (e.g., GpppG); dimethylated cap analog (e.g., m²′7GpppG), trimethylated cap analog (e.g., m^(2,2,7)GpppG), dimethylated symmetrical cap analogs (e.g., m⁷Gpppm⁷G), or anti reverse cap analogs (e.g., ARCA; m^(7,2,Ome)GpppG, m^(72,d)GpppG, m^(7,3,Ome)GpppG, m^(73,d)GpppG and their tetraphosphate derivatives) (see, e.g., Jemielity, J. et al., “Novel ‘anti-reverse’ cap analogs with superior translational properties”, RNA, 9: 1108-1122 (2003)).

In some embodiments, a suitable cap is a 7-methyl guanylate (“m⁷G”) linked via a triphosphate bridge to the 5′-end of the first transcribed nucleotide, resulting in m⁷G(5′)ppp(5′)N, where N is any nucleoside. A preferred embodiment of a m⁷G cap utilized in embodiments of the invention is m⁷G(5′)ppp(5′)G.

In some embodiments, the cap is a Cap0 structure. Cap0 structures lack a 2′-O-methyl residue of the ribose attached to bases 1 and 2. In some embodiments, the cap is a Cap1 structure. Cap1 structures have a 2′-O-methyl residue at base 2. In some embodiments, the cap is a Cap2 structure. Cap2 structures have a 2′-O-methyl residue attached to both bases 2 and 3.

A variety of m⁷G cap analogs are known in the art, many of which are commercially available. These include the m⁷GpppG described above, as well as the ARCA 3′-OCH₃ and 2′-OCH₃ cap analogs (Jemielity, J. et al., RNA, 9: 1108-1122 (2003)). Additional cap analogs for use in embodiments of the invention include N7-benzylated dinucleoside tetraphosphate analogs (described in Grudzien, E. et al., RNA, 10: 1479-1487 (2004)), phosphorothioate cap analogs (described in Grudzien-Nogalska, E., et al., RNA, 13: 1745-1755 (2007)), and cap analogs (including biotinylated cap analogs) described in U.S. Pat. Nos. 8,093,367 and 8,304,529, incorporated by reference herein.

Tail Structure

Typically, the presence of a “tail” serves to protect the mRNA from exonuclease degradation. The poly A tail is thought to stabilize natural messengers and synthetic sense RNA. Therefore, in certain embodiments a long poly A tail can be added to an mRNA molecule thus rendering the RNA more stable. Poly A tails can be added using a variety of art-recognized techniques. For example, long poly A tails can be added to synthetic or in vitro transcribed RNA using poly A polymerase (Yokoe, et al. Nature Biotechnology. 1996; 14: 1252-1256). A transcription vector can also encode long poly A tails. In addition, poly A tails can be added by transcription directly from PCR products. Poly A may also be ligated to the 3′ end of a sense RNA with RNA ligase (see, e.g., Molecular Cloning A Laboratory Manual, 2nd Ed., ed. by Sambrook, Fritsch and Maniatis (Cold Spring Harbor Laboratory Press: 1991 edition)).

In some embodiments, mRNAs (e.g., enzyme encoding mRNAs) include a 3′ poly(A) tail structure. Typically, the length of the poly A tail can be at least about 10, 50, 100, 200, 300, 400 at least 500 nucleotides. In some embodiments, a poly-A tail on the 3′ terminus of mRNA typically includes about 10 to 300 adenosine nucleotides (e.g., about 10 to 200 adenosine nucleotides, about 10 to 150 adenosine nucleotides, about 10 to 100 adenosine nucleotides, about 20 to 70 adenosine nucleotides, or about 20 to 60 adenosine nucleotides). In some embodiments, mRNAs include a 3′ poly(C) tail structure. A suitable poly-C tail on the 3′ terminus of mRNA typically include about 10 to 200 cytosine nucleotides (e.g., about 10 to 150 cytosine nucleotides, about 10 to 100 cytosine nucleotides, about 20 to 70 cytosine nucleotides, about 20 to 60 cytosine nucleotides, or about 10 to 40 cytosine nucleotides). The poly-C tail may be added to the poly-A tail or may substitute the poly-A tail.

In some embodiments, the length of the poly A or poly C tail is adjusted to control the stability of a modified sense mRNA molecule of the invention and, thus, the transcription of protein. For example, since the length of the poly A tail can influence the half-life of a sense mRNA molecule, the length of the poly A tail can be adjusted to modify the level of resistance of the mRNA to nucleases and thereby control the time course of polynucleotide expression and/or polypeptide production in a target cell.

5′ and 3′ Untranslated Region

In some embodiments, mRNAs include a 5′ and/or 3′ untranslated region. In some embodiments, a 5′ untranslated region includes one or more elements that affect an mRNA's stability or translation, for example, an iron responsive element. In some embodiments, a 5′ untranslated region may be between about 50 and 500 nucleotides in length.

In some embodiments, a 3′ untranslated region includes one or more of a polyadenylation signal, a binding site for proteins that affect an mRNA's stability of location in a cell, or one or more binding sites for miRNAs. In some embodiments, a 3′ untranslated region may be between 50 and 500 nucleotides in length or longer.

Exemplary 3′ and/or 5′ UTR sequences can be derived from mRNA molecules which are stable (e.g., globin, actin, GAPDH, tubulin, histone, or citric acid cycle enzymes) to increase the stability of the sense mRNA molecule. For example, a 5′ UTR sequence may include a partial sequence of a CMV immediate-early 1 (IE1) gene, or a fragment thereof to improve the nuclease resistance and/or improve the half-life of the polynucleotide. Also contemplated is the inclusion of a sequence encoding human growth hormone (hGH), or a fragment thereof to the 3′ end or untranslated region of the polynucleotide (e.g., mRNA) to further stabilize the polynucleotide. Generally, these modifications improve the stability and/or pharmacokinetic properties (e.g., half-life) of the polynucleotide relative to their unmodified counterparts, and include, for example modifications made to improve such polynucleotides' resistance to in vivo nuclease digestion.

According to various embodiments, any size mRNA may be encapsulated by provided liposomes. In some embodiments, the provided liposomes may encapsulate mRNA of greater than about 0.5 kb, 1 kb, 1.5 kb, 2 kb, 2.5 kb, 3 kb, 3.5 kb, 4 kb, 4.5 kb, or 5 kb in length.

Liposomes

The liposomes for use in provided compositions can be prepared by various techniques which are presently known in the art. For example, multilamellar vesicles (MLV) may be prepared according to conventional techniques, such as by depositing a selected lipid on the inside wall of a suitable container or vessel by dissolving the lipid in an appropriate solvent, and then evaporating the solvent to leave a thin film on the inside of the vessel or by spray drying. An aqueous phase may then added to the vessel with a vortexing motion which results in the formation of MLVs. Uni-lamellar vesicles (ULV) can then be formed by homogenization, sonication or extrusion of the multi-lamellar vesicles. In addition, unilamellar vesicles can be formed by detergent removal techniques.

In certain embodiments, provided compositions comprise a liposome wherein an agent, such as a nucleic acid e.g., mRNA, is associated on both the surface of the liposome and encapsulated within the same liposome. For example, during preparation of the compositions of the present invention, cationic liposomes may associate with the mRNA through electrostatic interactions. For example, during preparation of the compositions of the present invention, cationic liposomes may associate with the mRNA through electrostatic interactions.

In some embodiments, the compositions and methods of the invention comprise mRNA encapsulated in a liposome. In some embodiments, the one or more mRNA species may be encapsulated in the same liposome. In some embodiments, the one or more mRNA species may be encapsulated in different liposomes. In some embodiments, the mRNA is encapsulated in one or more liposomes, which differ in their lipid composition, molar ratio of lipid components, size, charge (Zeta potential), targeting ligands and/or combinations thereof. In some embodiments, the one or more liposome may have a different composition of cationic lipids, neutral lipid, PEG-modified lipid and/or combinations thereof. In some embodiments the one or more lipisomes may have a different molar ratio of cationic lipid, neutral lipid, cholesterol and PEG-modified lipid used to create the liposome.

The process of incorporation of a desired therapeutic agent, such as a nucleic acid (e.g., mRNA), into a liposome is often referred to as “loading”. Exemplary methods are described in Lasic, et al., FEBS Lett., 312: 255-258, 1992, which is incorporated herein by reference. The liposome-incorporated nucleic acids may be completely or partially located in the interior space of the liposome, within the bilayer membrane of the liposome, or associated with the exterior surface of the liposome membrane. The incorporation of a nucleic acid into liposomes is also referred to herein as “encapsulation” wherein the nucleic acid is entirely contained within the interior space of the liposome. The purpose of incorporating a mRNA into a transfer vehicle, such as a liposome, is often to protect the nucleic acid from an environment which may contain enzymes or chemicals that degrade nucleic acids and/or systems or receptors that cause the rapid excretion of the nucleic acids. Accordingly, in some embodiments, a suitable delivery vehicle is capable of enhancing the stability of the mRNA contained therein and/or facilitate the delivery of mRNA to the target cell or tissue.

Liposome Size

Suitable liposomes in accordance with the present invention may be made in various sizes. In some embodiments, provided liposomes may be made smaller than previously known mRNA encapsulating liposomes. In some embodiments, decreased size of liposomes is associated with more efficient delivery of mRNA. Selection of an appropriate liposome size may take into consideration the site of the target cell or tissue and to some extent the application for which the liposome is being made.

In some embodiments, an appropriate size of liposome is selected to facilitate systemic distribution of antibody encoded by the mRNA. In some embodiments, it may be desirable to limit transfection of the mRNA to certain cells or tissues. For example, to target hepatocytes a liposome may be sized such that its dimensions are smaller than the fenestrations of the endothelial layer lining hepatic sinusoids in the liver; in such cases the liposome could readily penetrate such endothelial fenestrations to reach the target hepatocytes.

Alternatively or additionally, a liposome may be sized such that the dimensions of the liposome are of a sufficient diameter to limit or expressly avoid distribution into certain cells or tissues. For example, a liposome may be sized such that its dimensions are larger than the fenestrations of the endothelial layer lining hepatic sinusoids to thereby limit distribution of the liposomes to hepatocytes.

In some embodiments, a suitable liposome has a size of or less than about 500 nm, 450 nm, 400 nm, 350 nm, 300 nm, 250 nm, 200 nm, 150 nm, 125 nm, 110 nm, 100 nm, 95 nm, 90 nm, 85 nm, 80 nm, 75 nm, 70 nm, 65 nm, 60 nm, 55 nm, or 50 nm. In some embodiments, a suitable liposome has a size no greater than about 250 nm (e.g., no greater than about 225 nm, 200 nm, 175 nm, 150 nm, 125 nm, 100 nm, 75 nm, or 50 nm). In some embodiments, a suitable liposome has a size ranging from about 10-250 nm (e.g., ranging from about 10-225 nm, 10-200 nm, 10-175 nm, 10-150 nm, 10-125 nm, 10-100 nm, 10-75 nm, or 10-50 nm). In some embodiments, a suitable liposome has a size ranging from about 100-250 nm (e.g., ranging from about 100-225 nm, 100-200 nm, 100-175 nm, 100-150 nm). In some embodiments, a suitable liposome has a size ranging from about 10-100 nm (e.g., ranging from about 10-90 nm, 10-80 nm, 10-70 nm, 10-60 nm, or 10-50 nm).

A variety of alternative methods known in the art are available for sizing of a population of liposomes. One such sizing method is described in U.S. Pat. No. 4,737,323, incorporated herein by reference. Sonicating a liposome suspension either by bath or probe sonication produces a progressive size reduction down to small ULV less than about 0.05 microns in diameter. Homogenization is another method that relies on shearing energy to fragment large liposomes into smaller ones. In a typical homogenization procedure, MLV are recirculated through a standard emulsion homogenizer until selected liposome sizes, typically between about 0.1 and 0.5 microns, are observed. The size of the liposomes may be determined by quasi-electric light scattering (QELS) as described in Bloomfield, Ann. Rev. Biophys. Bioeng., 10:421-150 (1981), incorporated herein by reference. Average liposome diameter may be reduced by sonication of formed liposomes. Intermittent sonication cycles may be alternated with QELS assessment to guide efficient liposome synthesis.

Pharmaceutical Compositions

To facilitate delivery of an agent, such as a nucleic acid e.g., mRNA, and/or expression of mRNA in vivo, delivery vehicles such as liposomes can be formulated in combination with one or more additional nucleic acids, carriers, targeting ligands or stabilizing reagents, or in pharmacological compositions where it is mixed with suitable excipients. Techniques for formulation and administration of drugs may be found in “Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton, Pa., latest edition.

Provided liposomally-encapsulated agents, such as a nucleic acid e.g., mRNA and compositions containing the same, may be administered and dosed in accordance with current medical practice, taking into account the clinical condition of the subject, the site and method of administration, the scheduling of administration, the subject's age, sex, body weight and other factors relevant to clinicians of ordinary skill in the art. The “effective amount” for the purposes herein may be determined by such relevant considerations as are known to those of ordinary skill in experimental clinical research, pharmacological, clinical and medical arts. In some embodiments, the amount administered is effective to achieve at least some stabilization, improvement or elimination of symptoms and other indicators as are selected as appropriate measures of disease progress, regression or improvement by those of skill in the art. For example, a suitable amount and dosing regimen is one that causes at least transient protein (e.g., enzyme) production.

Suitable routes of administration include, for example, oral, rectal, vaginal, transmucosal, pulmonary including intratracheal or inhaled, or intestinal administration; parenteral delivery, including intradermal, transdermal (topical), intramuscular, subcutaneous, intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, and/or intranasal administration.

Alternately or additionally, liposomally encapsulated agents, such as a nucleic acid e.g., mRNA and compositions of the invention may be administered in a local rather than systemic manner, for example, via injection of the pharmaceutical composition directly into a targeted tissue, preferably in a sustained release formulation. Local delivery can be affected in various ways, depending on the tissue to be targeted. For example, aerosols containing compositions of the present invention can be inhaled (for nasal, tracheal, or bronchial delivery); compositions of the present invention can be injected into the site of injury, disease manifestation, or pain, for example; compositions can be provided in lozenges for oral, tracheal, or esophageal application; can be supplied in liquid, tablet or capsule form for administration to the stomach or intestines, can be supplied in suppository form for rectal or vaginal application; or can even be delivered to the eye by use of creams, drops, or even injection. Formulations containing provided compositions complexed with therapeutic molecules or ligands can even be surgically administered, for example in association with a polymer or other structure or substance that can allow the compositions to diffuse from the site of implantation to surrounding cells. Alternatively, they can be applied surgically without the use of polymers or supports.

In some embodiments, provided liposomes and/or compositions are formulated such that they are suitable for extended-release of the agent, e.g., mRNA contained therein. Such extended-release compositions may be conveniently administered to a subject at extended dosing intervals. For example, in one embodiment, the compositions of the present invention are administered to a subject twice day, daily or every other day. In a preferred embodiment, the compositions of the present invention are administered to a subject twice a week, once a week, every ten days, every two weeks, every three weeks, or more preferably every four weeks, once a month, every six weeks, every eight weeks, every other month, every three months, every four months, every six months, every eight months, every nine months or annually. Also contemplated are compositions and liposomes which are formulated for depot administration (e.g., intramuscularly, subcutaneously, intravitreally) to either deliver or release a mRNA over extended periods of time. Preferably, the extended-release means employed are combined with modifications made to the mRNA to enhance stability.

Also contemplated herein are lyophilized pharmaceutical compositions comprising one or more of the liposomes disclosed herein and related methods for the use of such compositions as disclosed for example, in International Patent Application No. PCT/US2012/041663, filed Jun. 8, 2012, Publ. No. WO 2012/170889, the teachings of which are incorporated herein by reference in their entirety. For example, lyophilized pharmaceutical compositions according to the invention may be reconstituted prior to administration or can be reconstituted in vivo. For example, a lyophilized pharmaceutical composition can be formulated in an appropriate dosage form (e.g., an intradermal dosage form such as a disk, rod or membrane) and administered such that the dosage form is rehydrated over time in vivo by the individual's bodily fluids.

Provided liposomes and compositions may be administered to any desired tissue. In some embodiments, the agent, e.g., mRNA delivered by provided liposomes or compositions is expressed in the tissue in which the liposomes and/or compositions were administered. In some embodiments, the mRNA delivered is expressed in a tissue different from the tissue in which the liposomes and/or compositions were administered Exemplary tissues in which delivered mRNA may be delivered and/or expressed include, but are not limited to the liver, kidney, heart, spleen, serum, brain, skeletal muscle, lymph nodes, skin, and/or cerebrospinal fluid.

According to various embodiments, the timing of expression of delivered agents, e.g., mRNAs, can be tuned to suit a particular medical need. In some embodiments, the expression of the protein encoded by delivered mRNA is detectable 1, 2, 3, 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, and/or 72 hours in serum or target tissues after a single administration of provided liposomes or compositions. In some embodiments, the expression of the protein encoded by the mRNA is detectable 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, and/or 7 days in serum or target tissues after a single administration of provided liposomes or compositions. In some embodiments, the expression of the protein encoded by the mRNA is detectable 1 week, 2 weeks, 3 weeks, and/or 4 weeks in serum or target tissues after a single administration of provided liposomes or compositions. In some embodiments, the expression of the protein encoded by the mRNA is detectable after a month or longer after a single administration of provided liposomes or compositions.

Examples

While certain compounds, compositions and methods of the present invention have been described with specificity in accordance with certain embodiments, the following examples serve only to illustrate the compounds of the invention and are not intended to limit the same.

Example 1. Synthesis of Compounds of Formula I

A. Compounds of formula I, such as T23 (the compound of formula III), can be made according to the route shown in Scheme 1:

As shown in Scheme 1, exemplary compounds of formula I are prepared from L-lysine 1. L-lysine is first converted, via sodium nitrite and sulfuric acid, to a 6-amino-2-hydroxyhexanoic acid intermediate, which is treated with two equivalents of 2-decyloxirane to provide 6-(bis(2-hydroxydodecyl)amino)-2-hydroxyhexanoic acid 3, the alpha-hydroxy acid functionality of which is protected with 2,2-dimethoxypropane to yield 2,2-dimethyl-1,3-dioxolan-4-one 4. Protection of the secondary alcohols and deprotection of the alpha-hydroxy acid functionality of 5 yields 2-hydroxyhexanoic acid 6. 2-Hydroxyhexanoic acid 6 is split into two equivalent portions, one of which is converted to benzyl-protected alcohol 7 and the other is converted to Cbz-protected acid 8. Alcohol 7 and acid 8 are esterified using standard coupling protocols to yield ester 9. Subsequent hydrogenation to removal protecting groups affords the free acid 10, which was coupled intramolecularly to provide the lactone precursor 11. The final step required boron-mediated removal of TBS protecting groups yields the final target compound T23 (Compound of Formula III).

B. Similarly, T23 (the compound of formula III) can be made according to the route shown in Scheme 2:

As shown in Scheme 2, a short route to achieving T23 was achieved by exploiting direct intermolecular cyclization of the free a-hydroxy acid 6a to form the lactone precursor 11. Upon completion, identical boron-mediated removal of the TBS groups afforded the compound of Formula III, T23.

C. T23 can be also made according to the route shown in Scheme 3:

Synthesis of Compound 2.1

To a mixture of Boc-Lys-OH (18 g, 73 mmol) and DIPEA (16 mL) in methanol (216 mL) at room temperature was added (2S)-1,2-epoxydodecane (40 g, 219 mmol). The reaction mixture was heated at reflux overnight. The light yellow clear solution was concentrated to give a yellow oil, which was mixed with THF (120 mL), water (100 mL) and lithium hydroxide (6 g, 250 mmol) and stirred at room temperature overnight. The reaction mixture was then extracted with dichloromethane/methanol (9:1, 500 mL×4). The combined organic layers were dried over Na₂SO₄. Filtration and concentration gave 82 g crude product which was purified by flash chromatography on silica gel (1 kg, 0-35% methanol in ethyl acetate) to yield 38.6 g (81%) of 2.1 as an off-white solid.

Synthesis of Compound 3.1

A solution of compound 2.1 (38.6 g, 62.9 mmol) in anhydrous dichloromethane (200 mL) and trifluoroacetic acid (200 mL) was stirred at room temperature for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was dried under vacuum to give 39.5 g (100%) crude product 3.1 that was used directly for the next step without further purification.

Synthesis of Compound 4.1

To a mixture of compound 3.1 (39.5 g, 62.9 mmol) in 10% sulfuric acid (520 mL) at 0-5° C. (ice-water bath) with vigorously stirring, a solution of sodium nitrite (32 g, 464 mmol) in water (130 mL) was added dropwise in over 2 h while keeping the internal temperature below 5° C. After the addition was finished, the reaction mixture was allowed to warm up slowly to room temperature and stirred overnight. The reaction mixture was then extracted with dichloromethane/methanol (9:1, 800 mL×6). The combined organic layers were washed with saturated aqueous Na₂S₂O₃ solution and brine, then dried over Na₂SO₄. Filtration and concentration of the filtrate gave 38 g crude product which was purified using a Teledyne ISCO Combiflash automatic chromatography system (330 g Redisep silica gel column, 0-50% MeOH in CH₂Cl₂ gradient) to give 8.4 g of product 4.1 as light yellow foam (Yield: 73%, based on starting material consumed). 21 g of starting material 3.1 (free base) was also recovered.

Synthesis of Compound 5.1

Pyridinium p-toluenesulfonate (4.15 g, 16.5 mmol) was added to a solution of compound 4.1 (6.05 g, 11 mmol) in THF/2,2-dimethoxypropane (40 mL/40 mL). The resulting mixture was stirred at 55° C. for 5 h and 50° C. overnight. The solvents were removed under reduced pressure. The residue was dried under vacuum and used without purification.

Synthesis of Compound 6.1

The crude compound 5.1 was dissolved in DMF (30 mL). To this solution was added DMAP (269 mg, 2.2 mmol), imidazole (4.49 g, 66 mmol), and TBDMSCl (6.63 g, 44 mmol). The resulting solution was stirred at room temperature overnight. The solvents were removed under reduced pressure. The residue was partitioned between Et₂O (150 mL) and water (50 mL). The organic layer was separated, washed with brine (2×25 mL), dried over Na₂SO₄, and filtered. The filtrate was evaporated in vacuo and the residue was purified by column chromatography on silica gel (0-20% EtOAc/hexane) to give 5.4 g (63%, over two steps) of the desired product as a colorless oil.

Synthesis of Compound 7.1 (HCl Salt)

To a solution of compound 6.1 (5.32 g, 6.8 mmol) in THF (65 mL) was added dropwise 0.5 N NaOH (16.3 mL, 8.2 mmol). The resulting mixture was stirred at room temperature overnight. EtOAc (150 mL) was added. The mixture was acidified with 0.5 M HCl (40 mL). Then brine (60 mL) was added. The organic layer was separated, washed with brine (2×50 mL), dried over Na₂SO₄, and filtered. The filtrate was evaporated in vacuo. The residue was dried under vacuum to give 5.26 g (98%) of the desired product as an off-white wax.

Synthesis of Compound 7.1 (Sodium Salt)

To a solution of compound 6.1 (6.5 g, 8.3 mmol) in THF (80 mL) was added dropwise 0.5 N NaOH (20 mL, 10 mmol). The resulting mixture was stirred at room temperature overnight. Et₂O (200 mL) was added. The organic layer was washed with brine (3×50 mL), dried over Na₂SO₄, and filtered. The filtrate was evaporated in vacuo to give 6.5 g (99%) of the desired product as a colorless oil.

Synthesis of Compound 8.1

Benzyl bromide (1.08 mL, 9.1 mmol) was added dropwise to a solution of sodium salt of compound 7.1 (6.5 g, 8.3 mmol) in DMF/THF (30 mL/30 mL). The resulting solution was stirred at room temperature for 18 h. The solvents were removed under reduced pressure. The residue was taken up in EtOAc (150 mL). The organic layer was washed with water (25 mL), brine (2×25 mL), dried over Na₂SO₄, and filtered. The filtrate was evaporated in vacuo and the residue was purified by column chromatography on silica gel (0-20% EtOAc/hexane) to give 6.31 g (91%) of the desired product as a colorless oil.

Synthesis of Compound 9.1

To a cold (0° C.) solution of compound 7.1 HCl salt (5.26 g, 6.74 mmol) in THF/pyridine (15 mL/10 mL) was added dropwise benzyl chloroformate (1.15 mL, 8.1 mmol). The resulting mixture was allowed to warm to room temperature and stirred overnight. An aliquot of the reaction mixture was taken out for MS analysis. The result indicated the reaction did not go to completion. Benzyl chloroformate (1.15 mL, 8.1 mmol) was added. The reaction mixture was stirred at room temperature for another 1.5 h, then diluted with EtOAc (200 mL). The organic layer was washed with water (50 mL), 1.5 M HCl (2×50 mL), brine (3×40 mL), dried over Na₂SO₄, and filtered. The filtrate was evaporated in vacuo and the residue was purified by column chromatography on silica gel (0-70% EtOAc/hexane) to give 2.78 g (45%) of the desired product as light yellow oil.

Synthesis of Compound 10.1

To a cold (0° C.) solution of compound 8.1 (3.5 g, 4.2 mmol) and compound 9.1 (2.78 g, 3 mmol) in DCM (30 mL) was added DMAP (673 mg, 6 mmol) and DCC (2.48 g, 12 mmol). The resulting mixture was allowed to warm to room temperature and stirred overnight. DCM was removed under reduced pressure and the residue was taken up in Et₂O and filtered. The filtrate was evaporated in vacuo. The residue was purified by column chromatography on silica gel (0-20% EtOAc/hexane) to give 4.87 g (95%, contaminated with dicyclohexyl urea) of the desired product as a colorless oil.

Synthesis of Compound 11.1

To a solution of 10.1 (4.0 g, 2.36 mmol) in THF (anhydrous, 15 mL) in a 100 ml Teflon flask at 0° C. was added dropwise a 70% wt/30% wt HF-pyridine solution (15 mL, 578 mmol). The resulting mixture was stirred at room temperature for 2.5 h. Mass spectrometry analysis indicated completion of the reaction. The reaction solution was diluted with DCM (50 mL). The DCM solution was added to a mixture of DCM (200 mL) and aqueous Na₂CO₃ solution (40 g in 180 mL of water) with rapid stirring. The DCM layer was separated. The aqueous layer was extracted with DCM (150 mL). The combined organic phase was dried over Na₂SO₄ and evaporated. The light yellow oily residue was purified by silica gel column (120 g) on an ISCO automatic chromatography system eluting with 0-100% EtOAc in hexanes to give 2.66 g of 11.1 (77%) as a light yellow oil.

Synthesis of Compound 12.1

To a solution of 11.1 (2.66 g, 2.15 mmol) in THF (anhydrous, 70 mL) was added Et₃N (2.39 mL, 17.2 mmol), followed by TMSCl (1.49 mL, 11.8 mmol). The resulting mixture was stirred at room temperature overnight. Volatiles were removed. The residue was stirred with Et₂O (anhydrous, 100 mL) for 20 min and filtered. The solid was rinsed with Et₂O (anhydrous, 2×20 mL). The combined filtrate was evaporated and residue was dried under vacuum overnight to give 3.15 g of 12.1 (96%) as light yellow oil.

Synthesis of Compound 13.1

To a suspension of dry Pd/C (5%, 1.6 g) in EtOAc (15 mL) was added a solution of 12.1 (3.15 g, 2.1 mmol) in EtOAc (70 mL). The resulting mixture was stirred under a hydrogen balloon overnight. It was then filtered through Celite. The Celite was rinsed with EtOAc (25 mL×3). The combined filtrate was evaporated to give 2.09 g of 13.1 (78%) as a light yellow oil.

Synthesis of Target 23

To a solution of 13.1 (2.09 g, 1.6 mmol) in a mixture of DCM (anhydrous, 60 mL) and CH₃CN (anhydrous, 30 mL) was added DIPEA (0.415 mL, 2.4 mmol), followed by HATU (0.912 g, 2.4 mmol). The resulting mixture was stirred at room temperature under N₂ for 16 h. Volatiles were removed. The residue was extracted with hexane (100 mL+25 mL×2). The hexane extracts were combined and washed with aqueous NaHCO₃ (2×60 mL) and aq. HCl (5 mL 1 M HCl in 60 mL of H₂O). It was dried over Na₂SO₄ and filtered. The filtrate was evaporated to give compound 14.1 as light yellow foam which was dissolved in DCM (anhydrous, 25 mL). HCl in diethyl ether (2M, 6 mL) was added dropwise and the resulting mixture was stirred at room temperature under N₂ for 3 h. Mass spectrometry analysis indicated completion of the reaction. Solvents were removed by purging with a nitrogen gas flow. The residue was washed with anhydrous diethyl ether (10 mL×3) and dried under high vacuum to give 1.61 g of Target 23 as off-white solid (94%, two steps).

D. The compound of Formula IV, Target 24 (T24), can be made according to the route shown in Scheme 4:

Synthesis of Compound 3.2

A solution of propargylamine (4.83 g, 87.7 mmol) and 1,2-epoxydodecane (40.8 g, 210.4 mmol) in EtOH (300 mL) was heated under reflux for 16 h. The solvent was removed under reduced pressure and the residue was purified by column chromatography on silica gel (0-30% EtOAc/hexane) to give 32.8 g (88%) of the desired product as a light yellow solid.

Synthesis of Compound 4.2

To a cold (0° C.) solution of 3.2 (10.16 g, 24 mmol) in DMF (48 mL) was added sequentially DMAP (587 mg, 4.8 mmol), imidazole (5.72 g, 84 mmol), and tert-butyldimethylsilyl chloride (9.04 g, 60 mmol). The resulting mixture was stirred at 0° C. for 20 min. The ice bath was then removed, and the reaction was allowed to warm to room temperature and stirred overnight. DMF was removed under reduced pressure. To the residue was added EtOAc (200 mL), water (50 mL), and brine (30 mL). The organic layer was separated, washed with brine (50 mL), dried over Na₂SO₄, and filtered. The filtrate was evaporated in vacuo and the residue was purified by column chromatography on silica gel (hexane) to give 14.55 g (93%) of the desired product as colorless oil.

Synthesis of Compound 6.2

To a cold (0° C.) solution of 4.2 (3.52 g, 5.4 mmol) in toluene (40 mL) was added dropwise 2.5 M n-BuLi (2.16 mL, 5.4 mmol). The resulting solution was stirred at 0° C. for 30 min, then 1 M Et₂AlCl (5.4 mL, 5.4 mmol) was added dropwise. After addition, the cloudy solution was stirred at 0° C. for another 2 h, then a solution of 5.2 (375 mg, 2.88 mmol) in toluene (2 mL) was added dropwise. After stirring for 30 min, the ice bath was removed, and the reaction was allowed to warm up to room temperature and stirred overnight. The reaction was cooled with an ice bath then Na₂SO₄.10H₂O (8.3 g) was added in one portion. The resulting mixture was stirred vigorously for 2 h. The mixture was filtered and the filtrate was evaporated in vacuo. The residue was purified by column chromatography on silica gel (0-5% EtOAc/hexane) twice to give 1.12 g (53%) of the desired product as a yellow oil.

Synthesis of Compound 7.2

To a solution of 6.2 (2.0 g, 2.56 mmol) in EtOH (50 mL) was added pyridine (1.45 mL, 17.9 mmol) and 5 wt. % Pd/C (272 mg, 0.128 mmol). The resulting mixture was degassed with Ar three times and then stirred under 1 atm H₂ overnight. The mixture was filtered through a Celite plug which was washed with EtOH thoroughly. The combined filtrate and washes were evaporated in vacuo to give 2.0 g of light yellow oil as a mixture of the desired product and other inseparable byproducts. The crude was used in next step without further purification.

Synthesis of Compound 8.2

To a solution of crude 7.2 (2.0 g) in THF (31 mL) was added 0.5 N NaOH (6.14 mL, 3.07 mmol) and MeOH (1.5 mL). The resulting mixture was stirred vigorously at room temperature overnight. The solvent was removed under reduced pressure. The residue was taken up in Et₂O (120 mL), and was washed with brine (3×30 mL), dried over Na₂SO₄, and filtered. The residue was purified by column chromatography on silica gel (0-50% EtOAc/hexane) to give 739 mg (37% over two steps) of the desired product as a light yellow oil.

Synthesis of Compound 9.2

To a solution of compound 8.2 (739 mg, 0.97 mmol) in DMF/THF (5 mL/10 mL) at room temperature was added K₂CO₃ (201 mg, 1.46 mmol). The resulting mixture was stirred for 20 min then benzyl bromide (127 μL, 1.07 mmol) was added. The resulting mixture was stirred at room temperature overnight. The solvents were removed under reduced pressure. The residue was taken up in EtOAc (70 mL) and washed with water (10 mL), brine (15 mL), dried over Na₂SO₄, and filtered. The filtrate was evaporated in vacuo and the residue was purified by column chromatography on silica gel (0-10% EtOAc/hexane) to give 796 mg (97%) of the desired product as colorless oil.

Synthesis of Compound 10.2

To a cold (0° C.) solution of 8.2 (795 mg, 1.05 mmol) in THF (10.5 mL) was added sequentially 2,6-lutidine (128 μL, 1.1 mmol) and benzyl chloroformate (157 μL, 1.1 mmol). After stirring for 15 min, the ice bath was removed. The reaction was allowed to warm to room temperature and stirred for 4 h. An aliquot of reaction mixture was taken out for MS analysis. The result indicated the reaction did not go to completion. Then benzyl chloroformate (157 μL, 1.1 mmol) was added. After stirring at room temperature for another 1 h, the reaction was quenched with saturated aqueous NaHCO₃ solution (25 mL). The mixture was stirred vigorously overnight. The mixture was diluted with EtOAc (70 mL). The organic layer was separated and the aqueous layer was extracted with EtOAc (20 mL). The combined organic layers were washed with 0.5 M HCl (20 mL), brine (2×15 mL), dried over Na₂SO₄, and filtered. The filtrate was evaporated in vacuo and the residue was purified by column chromatography on silica gel (0-50% EtOAc/hexane) to give 663 mg (67%) of the desired product as a yellow oil.

Synthesis of Compound 11.2

To a cold (0° C.) solution of compound 9.2 (800 mg, 0.94 mmol) and 10.2 (664 mg, 0.71 mmol) in CH₂Cl₂ (7.1 mL) was added DMAP (157 mg, 1.42 mmol) and DCC (293 mg, 1.42 mmol). After stirring for 15 min, the reaction was allowed to warm up to room temperature and stirred overnight. CH₂Cl₂ was removed under reduced pressure. The residue was taken up in Et₂O (50 mL). The white solid was removed by filtration. The filtrate was evaporated in vacuo. The residue was purified by column chromatography on silica gel (0-10% EtOAc/hexane) twice to give 964 mg (79%) of product 11.2 as a colorless oil.

Synthesis of Compound 12.2

To a solution of 11.2 (5.04 g, 2.93 mmol) in THF (anhydrous, 15 mL) was added the 70% wt/30% wt HF-pyridine solution (20 mL, 770 mmol). The resulting mixture was stirred at room temperature. Mass spectrometry analysis after 2.5 h indicated complete reaction. THF was removed. The residual solution was diluted with DCM (50 mL). The DCM solution was added to a mixture of DCM (200 mL) and aqueous Na₂CO₃ solution (61 g in 300 mL of water) with rapid stirring. The DCM layer was separated. The aqueous layer was extracted with DCM (150 mL). The combined organic phase was dried over Na₂SO₄ and evaporated. The light yellow oily residue was purified by silica gel column (80 g) on an ISCO automatic chromatography system eluting with 0-100% EtOAc in hexane to give 2.85 g of 12.2 (77%) as a light yellow oil.

Synthesis of Compound 13.2

To a solution of 12.2 (2.85 g, 2.25 mmol) in THF (anhydrous, 70 mL) was added Et₃N (2.5 mL, 18 mmol), followed by TMSCl (1.5 mL, 11.9 mmol). The resulting mixture was stirred at room temperature for 3 h. Volatiles were removed. The residue was stirred with Et₂O (anhydrous, 100 mL) for 20 min and filtered. The solid was rinsed with Et₂O (anhydrous, 2×20 mL). The combined filtrate was evaporated and residue was dried under vacuum overnight to give 3.29 g of 13.2 (94%) as a light yellow oil.

Synthesis of Compound 14.2

To a suspension of Pd/C (5%, 1.62 g, mmol) in EtOAc (10 mL) was added a solution of 13.2 (3.29 g, 2.11 mmol) in EtOAc (70 mL). The resulting mixture was stirred under a balloon of H₂ for 2 h. It was then filtered through Celite. The Celite was rinsed with EtOAc (3×20 mL). The combined filtrate was evaporated to give 2.78 g of 14.2 (99%) as a light yellow oil.

Synthesis of Compound 15.2

To a solution of 14.2 (2.78 g, 2.09 mmol) in a mixture of DCM (anhydrous, 40 mL) and CH₃CN (anhydrous, 20 mL) was added DIPEA (0.55 mL, 3.14 mmol), followed by HATU (1.19 g, 3.14 mmol). The resulting mixture was stirred at room temperature under N₂ for 16 h. Volatiles were removed. The residue was extracted with hexane (150 mL). The hexane extract was washed with aqueous NaHCO₃ (2×60 mL) and HCl. It was dried over Na₂SO₄ and filtered. The filtrate was evaporated to give 2.36 g (86%) of 15.2 as a light yellow gum. Synthesis of Target 24

To a solution of 15.2 (1.09 g, 0.83 mmol) in DCM (anhydrous, 15 mL), HCl in diethyl ether (2 M, 3 mL) was added dropwise and the resulting mixture was stirred at room temperature under N₂ for 3.5 h. Mass spectrometry analysis indicated completion of the reaction. Solvent was removed by purging with a nitrogen gas flow. The residue was washed with anhydrous diethyl ether (20 mL×3) and dried under high vacuum to give 875 mg of crude Target 24. 255 mg of crude Target 24 was washed with anhydrous acetonitrile (30 mL×3). The residue was dissolved in DCM (anhydrous, 2 mL) and added to a mixture of diethyl ether (anhydrous, 25 mL) and HCl in diethyl ether (2 M, 0.5 mL) with stirring. After continued stirring for 30 minutes, the gummy solid was separated from the solution and was washed with anhydrous diethyl ether (5 mL×2). It was dried under high vacuum to give 200 mg (69%) of Target 24 as an off-white foam.

Example 2. Exemplary Liposome Formulations for mRNA Delivery and Expression

This example provides exemplary liposome formulations incorporating the cationic lipids described in this application, for example, the compound of formula III, for effective delivery and expression of mRNA encoding therapeutic proteins in vivo. Lipid Materials

In general, the formulations described herein are based on a multi-component lipid mixture of varying ratios employing one or more cationic lipids, one or more helper lipids (e.g., non-cationic lipids and/or cholesterol-based lipids), and one or more PEGylated lipids designed to encapsulate various nucleic acid-based materials. As a non-limiting example, the compound of Formula III (3,6-bis(4-(bis(2-hydroxydodecyl)amino)butyl)-1,4-dioxane-2,5-dione) is used in various formulations described herein. Exemplary helper lipids include one or more of DSPC (1,2-distearoyl-sn-glycero-3-phosphocholine), DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine), DOPE (1,2-dioleyl-sn-glycero-3-phosphoethanolamine), DOPC (1,2-dioleyl-sn-glycero-3-phosphotidylcholine) DPPE (1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine), DMPE (1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine), DOPG (1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol)), cholesterol, etc. Exemplary PEGylated lipids include a poly(ethylene) glycol chain of up to 5 kDa in length covalently attached to a lipid with alkyl chain(s) of C₆-C₂₀ length, for example, PEG-2K. As non-limiting examples, liposome formulations used in various examples described herein include the compound of Formula III, DOPE, cholesterol and DMG-PEG2K at various ratios. For example, in some cases, the ratio of the compound of Formula III:DOPE:cholesterol:DMG-PEG2K is approximately 40:30:20:10 by weight. In other cases, the ratio of the compound of Formula III:DOPE:cholesterol:DMG-PEG2K is approximately 40:32:25:3 by weight. Unless otherwise specified, the below Examples include a mixture in the ratio of the compound of Formula III:DOPE:cholesterol:DMG-PEG2K of approximately 40:30:25:5 by weight.

Messenger RNA Material

The formulations described herein may be used to deliver any mRNA, in particular, therapeutic mRNA. As used herein, a therapeutic mRNA refers to an mRNA that encodes a therapeutic protein. The formulations described herein can also be used to deliver any modified or unmodified mRNA, or mRNA with naturally occurring sequences or codon-optimized.

As non-limiting examples, human Factor IX (FIX), codon-optimized Firefly Luciferase (FFL), codon-optimized human argininosuccinate synthetase (ASS1) messenger RNA, codon-optimized human Spinal Motor Neuron 1(SMN) mRNA were synthesized by in vitro transcription from a plasmid DNA template encoding the gene, which was followed by the addition of a 5′ cap structure (Cap 1) (Fechter, P.; Brownlee, G. G. “Recognition of mRNA cap structures by viral and cellular proteins” J. Gen. Virology 2005, 86, 1239-1249) and a 3′ poly(A) tail of, e.g., approximately 250 nucleotides in length as determined by gel electrophoresis. Typically, 5′ and 3′ untranslated regions (UTR) are present in each mRNA product and are represented as X and Y, respectively. Example 5′ and 3′ UTR sequences are described below. The exemplary sequences of FIX, ASS1, and FFL mRNA used in the examples herein are listed below. Also shown are the 5′ and 3′ UTR sequences.

Codon-Optimnized Firefly Luciferase (FFL) mRNA.

(SEQ ID NO.: 3) XAUGGAAGAUGCCAAAAACAUUAAGAAGGGCCCAGCGCCAUUCUACCCAC UCGAAGACGGGACCGCCGGCGAGCAGCUGCACAAAGCCAUGAAGCGCUAC GCCCUGGUGCCCGGCACCAUCGCCUUUACCGACGCACAUAUCGAGGUGGA CAUUACCUACGCCGAGUACUUCGAGAUGAGCGUUCGGCUGGCAGAAGCUA UGAAGCGCUAUGGGCUGAAUACAAACCAUCGGAUCGUGGUGUGCAGCGAG AAUAGCUUGCAGUUCUUCAUGCCCGUGUUGGGUGCCCUGUUCAUCGGUGU GGCUGUGGCCCCAGCUAACGACAUCUACAACGAGCGCGAGCUGCUGAACA GCAUGGGCAUCAGCCAGCCCACCGUCGUAUUCGUGAGCAAGAAAGGGCUG CAAAAGAUCCUCAACGUGCAAAAGAAGCUACCGAUCAUACAAAAGAUCAU CAUCAUGGAUAGCAAGACCGACUACCAGGGCUUCCAAAGCAUGUACACCU UCGUGACUUCCCAUUUGCCACCCGGCUUCAACGAGUACGACUUCGUGCCC GAGAGCUUCGACCGGGACAAAACCAUCGCCCUGAUCAUGAACAGUAGUGG CAGUACCGGAUUGCCCAAGGGCGUAGCCCUACCGCACCGCACCGCUUGUG UCCGAUUCAGUCAUGCCCGCGACCCCAUCUUCGGCAACCAGAUCAUCCCC GACACCGCUAUCCUCAGCGUGGUGCCAUUUCACCACGGCUUCGGCAUGUU CACCACGCUGGGCUACUUGAUCUGCGGCUUUCGGGUCGUGCUCAUGUACC GCUUCGAGGAGGAGCUAUUCUUGCGCAGCUUGCAAGACUAUAAGAUUCAA UCUGCCCUGCUGGUGCCCACACUAUUUAGCUUCUUCGCUAAGAGCACUCU CAUCGACAAGUACGACCUAAGCAACUUGCACGAGAUCGCCAGCGGCGGGG CGCCGCUCAGCAAGGAGGUAGGUGAGGCCGUGGCCAAACGCUUCCACCUA CCAGGCAUCCGCCAGGGCUACGGCCUGACAGAAACAACCAGCGCCAUUCU GAUCACCCCCGAAGGGGACGACAAGCCUGGCGCAGUAGGCAAGGUGGUGC CCUUCUUCGAGGCUAAGGUGGUGGACUUGGACACCGGUAAGACACUGGGU GUGAACCAGCGCGGCGAGCUGUGCGUCCGUGGCCCCAUGAUCAUGAGCGG CUACGUUAACAACCCCGAGGCUACAAACGCUCUCAUCGACAAGGACGGCU GGCUGCACAGCGGCGACAUCGCCUACUGGGACGAGGACGAGCACUUCUUC AUCGUGGACCGGCUGAAGAGCCUGAUCAAAUACAAGGGCUACCAGGUAGC CCCAGCCGAACUGGAGAGCAUCCUGCUGCAACACCCCAACAUCUUCGACG CCGGGGUCGCCGGCCUGCCCGACGACGAUGCCGGCGAGCUGCCCGCCGCA GUCGUCGUGCUGGAACACGGUAAAACCAUGACCGAGAAGGAGAUCGUGGA CUAUGUGGCCAGCCAGGUUACAACCGCCAAGAAGCUGCGCGGUGGUGUUG UGUUCGUGGACGAGGUGCCUAAAGGACUGACCGGCAAGUUGGACGCCCGC AAGAUCCGCGAGAUUCUCAUUAAGGCCAAGAAGGGCGGCAAGAUC GCCGUGUAAY  5′ and 3′ UTR Sequences X (5′ UTR Sequence) = (SEQ ID NO.: 5) GGACAGAUCGCCUGGAGACGCCAUCCACGCUGUUUUGACCUCCAUAGAAG ACACCGGGACCGAUCCAGCCUCCGCGGCCGGGAACGGUGCAUUGGAACGC GGAUUCCCCGUGCCAAGAGUGACUCACCGUCCUUGACACG  Y (3′ UTR Sequence) = (SEQ ID NO.: 6) CGGGUGGCAUCCCUGUGACCCCUCCCCAGUGCCUCUCCUGGCCCUGGAA GUUGCCACUCCAGUGCCCACCAGCCUUGUCCUAAUAAAAUUAAGUUGCA UCAAGCU  Or (SEQ ID NO.: 7) GGGUGGCAUCCCUGUGACCCCUCCCCAGUGCCUCUCCUGGCCCUGGAAG UUGCCACUCCAGUGCCCACCAGCCUUGUCCUAAUAAAAUUAAGUUGCAU CAAGCU 

Aliquots of 50 mg/mL ethanolic solutions of the compound of Formula III, DOPE, Chol and DMG-PEG2K are mixed in a molar ratio of 40:30:25:5 and diluted with ethanol to 3 mL final volume. Separately, an aqueous buffered solution (10 mM citrate/150 mM NaCl, pH 4.5) of FIX, ASS1, or FFL mRNA is prepared from a 1 mg/mL stock. The lipid solution is injected rapidly into the aqueous mRNA solution and shaken to yield a final suspension in 20% ethanol. The resulting nanoparticle suspension is filtered, diafiltrated with 1×PBS (pH 7.4), concentrated and stored at 2-8° C. The final concentration of FIX mRNA is typically diluted to approximately 0.20 mg/mL FIX mRNA (encapsulated), Z_(ave)=76 nm, PDI=0.08. The final concentration of ASS1 mRNA is typically diluted to approximately 0.20 mg/mL ASS1 mRNA (encapsulated), Z_(ave)=78 nm (Dv(50)=46 nm; Dv(90)=96 nm). The final concentration of FFL mRNA is is typically diluted to approximately 0.20 mg/mL FFL mRNA (encapsulated), Z_(ave)=75 nm, PDI −0.11. The final concentration of SMN mRNA is is typically diluted to approximately 0.20 mg/mL SMN mRNA (encapsulated). Average particle size (Z_(ave))=71 nm, (particle size for 50% of particles was 44 nm or less (Dv(50))=44 nm; and the particle size for 90% of the particles was 93n or less (Dv(90)=93 nm)).

Exemplary Formulation Comprising T23

Aliquots of 50 mg/mL ethanolic solutions of the compound of Target 23, DOPE, Chol and DMG-PEG2K were mixed in a molar ratio of 40:30:25:5 and diluted with ethanol to 3 mL final volume. Separately, an aqueous buffered solution (10 mM citrate/150 mM NaCl, pH 4.5) of EPO mRNA was prepared from a 1 mg/mL stock. The lipid solution was injected rapidly into the aqueous mRNA solution and shaken to yield a final suspension in 20% ethanol. The resulting nanoparticle suspension was diafiltrated with 1×PBS (pH 7.4), concentrated and stored at 2-8° C. The final concentration of EPO mRNA is typically diluted to approximately 0.20 mg/mL EPO mRNA (encapsulated). Z_(ave)=80 nm, PDI=0.11.

Exemplary Formulation Comprising T24

Aliquots of 50 mg/mL ethanolic solutions of the compound of Target 24, DOPE, Chol and DMG-PEG2K were mixed in a molar ratio of 40:30:25:5 and diluted with ethanol to 3 mL final volume. Separately, an aqueous buffered solution (10 mM citrate/150 mM NaCl, pH 4.5) of EPO mRNA was prepared from a 1 mg/mL stock. The lipid solution was injected rapidly into the aqueous mRNA solution and shaken to yield a final suspension in 20% ethanol. The resulting nanoparticle suspension was diafiltrated with 1×PBS (pH 7.4), concentrated and stored at 2-8° C. The final concentration of EPO mRNA is typically diluted to approximately 0.20 mg/mL EPO mRNA (encapsulated). Z_(ave)=78 nm, PDI=0.14.

Example 3. In Vivo Results

CD-1 Mice (N=4 per group) were injected with a 0.20 mg/mL formulation of either Target 23-based LNPs or Target 24-based LNPs loaded with hEPO mRNA (1.0 mg/kg). Serum levels of hEPO were monitored at 6 hr and 24 hr post-dose. See FIG. 1. Liver enzymes (ALT/AST) were measured 24 hr post-administration. See Table 1.

TABLE 1 Liver enzymes levels in wild type mouse sera after treatment via hEPO mRNA loaded LNPs. Formulation ALT Levels AST Levels Target 23 LNP 107 ± 37 92 ± 17 Target 24 LNP  77 ± 16 73 ± 11

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. The scope of the present invention is not intended to be limited to the above Description, but rather is as set forth in the following claims: 

We claim:
 1. A compound according to formula I:

or a pharmaceutically acceptable salt thereof, wherein: each instance of X is independently O or S; each instance of Y is independently O or S; each instance of R¹ is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, halogen, —OR^(A1), —N(R^(A1))₂, —SR^(A1), or a group of formula (iv):

L is an optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted heteroalkenylene, optionally substituted heteroalkynylene, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene, or combination thereof, and each of R⁶ and R⁷ is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group, or a group of formula (i), (ii) or (iii); formulae (i), (ii), and (iii) are:

each instance of R′ is independently hydrogen or optionally substituted alkyl; X′ is O or S, or NR^(X); R^(X) is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or a nitrogen protecting group; Y′ is O, S, or NR^(Y); R^(Y) is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or a nitrogen protecting group; R^(P) is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, an oxygen protecting group when attached to an oxygen atom, a sulfur protecting group when attached to a sulfur atom, or a nitrogen protecting group when attached to a nitrogen atom; R^(L) is optionally substituted C₁₋₅₀ alkyl, optionally substituted C₂₋₅₀ alkenyl, optionally substituted C₂₋₅₀ alkynyl, optionally substituted heteroC₁₋₅₀ alkyl, optionally substituted heteroC₂₋₅₀ alkenyl, optionally substituted heteroC₂₋₅₀ alkynyl, or a polymer; and each occurrence of R^(A1) is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, an oxygen protecting group when attached to an oxygen atom, a sulfur protecting group when attached to an sulfur atom, a nitrogen protecting group when attached to a nitrogen atom, or two R^(A1) groups, together with the nitrogen atom to which they are attached, are joined to form an optionally substituted heterocyclic or optionally substituted heteroaryl ring.
 2. The compound of claim 1, wherein the compound is of formula II:

or a pharmaceutically acceptable salt thereof, wherein: each X independently is O or S; each Y independently is O or S; each m independently is 0 to 20; each n independently is 1 to 6; each R_(A) is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl or halogen; and each R_(B) is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl or halogen.
 3. The compound of claim 2, wherein the compound is of formula III:

or a pharmaceutically acceptable salt thereof.
 4. The compound of claim 2 wherein the compound is of formula IV:

or a pharmaceutically acceptable salt thereof.
 5. The compound of claim 2 wherein the compound is of formula V:

or a pharmaceutically acceptable salt thereof.
 6. A composition for delivery of a polynucleotide comprising a compound of any one of the preceding claims.
 7. The composition of claim 6, wherein the composition is a liposome.
 8. The composition of claim 7, wherein the liposome further comprises one or more non-cationic lipids, one or more cholesterol-based lipids and/or one or more PEG-modified lipids.
 9. The composition of claim 8, wherein the one or more non-cationic lipids are selected from DSPC (1,2-distearoyl-sn-glycero-3-phosphocholine), DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine), DOPE (1,2-dioleyl-sn-glycero-3-phosphoethanolamine), DOPC (1,2-dioleyl-sn-glycero-3-phosphotidylcholine) DPPE (1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine), DMPE (1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine), DOPG (1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol)).
 10. The composition of claim 8, wherein the one or more PEG-modified lipids comprise a poly(ethylene) glycol chain of up to 5 kDa in length covalently attached to a lipid with alkyl chain(s) of C₆-C₂₀ length.
 11. The composition of any one of claims 7-10, wherein the liposome has a size less than about 250 nm, 200 nm, 150 nm, 100 nm, 75 nm, or 50 nm.
 12. A method of delivery of messenger RNA (mRNA) in vivo, comprising administering to a subject in need of delivery a composition comprising an mRNA encoding a protein, encapsulated within a liposome such that the administering of the composition results in the expression of the protein encoded by the mRNA in vivo; wherein the liposome comprises a cationic lipid compound of any one of claims 1-5.
 13. The method of claim 12, wherein the cationic lipid is of formula II:

or a pharmaceutically acceptable salt thereof, wherein: each X independently is O or S; each Y independently is O or S; each m independently is 0 to 20; each n independently is 1 to 6; each R_(A) is independently hydrogen, optionally substituted C1-50 alkyl, optionally substituted C2-50 alkenyl, optionally substituted C2-50 alkynyl, optionally substituted C3-10 carbocyclyl, optionally substituted 3-14 membered heterocyclyl, optionally substituted C6-14 aryl, optionally substituted 5-14 membered heteroaryl or halogen; and each R_(B) is independently hydrogen, optionally substituted C1-50 alkyl, optionally substituted C2-50 alkenyl, optionally substituted C2-50 alkynyl, optionally substituted C3-10 carbocyclyl, optionally substituted 3-14 membered heterocyclyl, optionally substituted C6-14 aryl, optionally substituted 5-14 membered heteroaryl or halogen.
 14. The method of claim 12, wherein the cationic lipid is of formula III:

or a pharmaceutically acceptable salt thereof.
 15. The method of claim 12 wherein the compound is of formula IV:

or a pharmaceutically acceptable salt thereof.
 16. The method of claim 12 wherein the compound is of formula V:

or a pharmaceutically acceptable salt thereof.
 17. The method of any one of claims 12-16, wherein the liposome further comprises one or more non-cationic lipids, one or more cholesterol-based lipids and/or one or more PEG-modified lipids.
 18. The method of claim 17, wherein the one or more non-cationic lipids are selected from DSPC (1,2-distearoyl-sn-glycero-3-phosphocholine), DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine), DOPE (1,2-dioleyl-sn-glycero-3-phosphoethanolamine), DOPC (1,2-dioleyl-sn-glycero-3-phosphotidylcholine) DPPE (1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine), DMPE (1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine), DOPG (1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol)).
 19. The method of claim 17 or 18, wherein the one or more cholesterol-based lipids are cholesterol and/or PEGylated cholesterol.
 20. The method of any one of claims 17-19, wherein the one or more PEG-modified lipids comprise a poly(ethylene) glycol chain of up to 5 kDa in length covalently attached to a lipid with alkyl chain(s) of C₆-C₂₀ length.
 21. The method of any one of claims 12-20, wherein the liposome has a size less than about 250 nm, 200 nm, 150 nm, 100 nm, 75 nm, or 50 nm.
 22. The method of any one of claims 12-21, wherein the mRNA has a length of or greater than about 0.5 kb, 1 kb, 1.5 kb, 2 kb, 2.5 kb, 3 kb, 3.5 kb, 4 kb, 4.5 kb, or 5 kb.
 23. The method of any one of claims 12-22, wherein the protein encoded by the mRNA is a cytosolic protein.
 24. The method of any one of claims 12-22, wherein the protein encoded by the mRNA is a secreted protein.
 25. The method of any one of claims 12-22, wherein the protein encoded by the mRNA is an enzyme.
 26. The method of any one of claims 12-25, wherein the mRNA comprises one or more modified nucleotides.
 27. The method of claim 26, wherein the one or more modified nucleotides comprise pseudouridine, N-1-methyl-pseudouridine, 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deaza-guanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, and/or 2-thiocytidine.
 28. The method of any one of claims 12-27, wherein the mRNA is unmodified.
 29. A method of treating a disease or disorder comprising a step of delivering an mRNA encoding a therapeutic protein using a method of any of claims 12-28. 